From 340f3b20ca2ffb5370efbe6567e88b32bb3a86e1 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Wed, 8 Jul 2026 14:05:20 +0100 Subject: [PATCH 01/47] polish contributions.md --- docs/CONTRIBUTING.md | 12 ++++-------- 1 file changed, 4 insertions(+), 8 deletions(-) diff --git a/docs/CONTRIBUTING.md b/docs/CONTRIBUTING.md index 67a4370..94bbd9f 100644 --- a/docs/CONTRIBUTING.md +++ b/docs/CONTRIBUTING.md @@ -204,13 +204,11 @@ When adding new functionality (e.g. a new submission mode such as `isolate_genom All inputs in seqsubmit are defined via strict samplesheet schemas. -- Any new feature must: - - Extend or introduce a corresponding schema (assets/schema*input*\*.json) - - Clearly define required and optional fields - Maintain: - Column order consistency (critical for pipeline execution) - Backward compatibility where possible - Avoid duplicating metadata logic—reuse existing parsing and validation patterns. +- New feature should extend or introduce a corresponding schema (`assets/schema_\*.json`) if pipeline inputs are affected. ⸻ @@ -220,8 +218,6 @@ Submissions to [ENA](https://www.ebi.ac.uk/ena/browser/home) are governed by a w Contributors should refer directly to the [official ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/general-guide.html) to understand latest submission requirements and ensure that pipeline behaviour remains aligned with ENA standards. - Follow ENA-defined submission workflows and required steps for each data type - - Ensure that: - New pipeline workflows follow ENA-defined submission workflows and required steps for each data type - Metadata fields align with ENA requirements @@ -237,8 +233,8 @@ Contributors should refer directly to the [official ENA documentation](https://e All submissions are performed via [Webin-CLI](https://github.com/enasequence/webin-cli). Contributors should rely on official Webin-CLI documentation to understand expected inputs, parameters, and modes of operation. Make sure to always use the latest version of the tool. - Contributions affecting submission logic must: - - Respect --webincli_mode (validate vs submit) - - Support --test_upload for safe testing against ENA test server + - Respect `--webincli_mode` (validate vs submit) + - Support `--test_upload` for safe testing against ENA test server - Never introduce changes that: - Risk accidental submission to production without explicit user intent - Ensure compatibility with: @@ -276,7 +272,7 @@ Some data processing steps may be resource-intensive (e.g. database preparation 7. Documentation and Usability -Every contribution should improve usability: +Every contribution should aim to improve usability: - Update: - README.md From 9692ab3827dea6abb994b3361ecd125892a037f9 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Wed, 8 Jul 2026 14:38:08 +0100 Subject: [PATCH 02/47] add limitations block --- README.md | 20 ++++++++++++++++++++ 1 file changed, 20 insertions(+) diff --git a/README.md b/README.md index 40f2f6e..7133ca1 100644 --- a/README.md +++ b/README.md @@ -31,6 +31,26 @@ Currently, the pipeline supports four submission modes, each routed to a dedicat ![seqsubmit workflow diagram](assets/seqsubmit_schema.png) +## Limitations + +nf-core/seqsubmit does not yet support the following data types and scenarios: + +- **Co-assemblies**: Metagenomic co-assemblies, and MAGs/bins generated from co-assemblies, are not supported. The pipeline currently expects each assembly/MAG/bin to be derived from a single run or an assembly generated from a single run. + +- **Eukaryotic and viral bins/MAGs**: Submitting eukaryotic or viral genomes that require additional metadata generation is not supported. + - For **eukaryotic bins/MAGs**, RNA prediction and quality assessment (completeness/contamination estimation) are not implemented. + - For **viral bins/MAGs**, RNA prediction, quality assessment (completeness/contamination estimation) and taxonomy assignment are not implemented. + +- **Reads in non-FASTQ formats**: Only reads provided as FASTQ (`.fastq.gz`) can be submitted with `--mode reads`. Other formats (e.g. BAM) are not supported as input. + +- **Submission of MAG/bin without submitting the assembly or raw reads**: If a MAG/bin cannot be traced back to an assembly accession or a reads accession — i.e. only a sample accession is available — it cannot currently be submitted, since the pipeline relies on that lineage to build the required metadata. + +- **Submission of assembly without submitting the raw reads**: Similarly, assemblies that have no associated reads accession (`run_accession` column in the sample sheet) are not supported for the same reason. + +- **Single-contig assemblies/MAGs/bins**: Assemblies or MAGs/bins that consist of only one contig cannot be uploaded through this pipeline. ENA classifies these as "chromosomal assemblies", which follow a different submission procedure with different metadata requirements. If you have single-contig assembly/MAG/bin to submit, please contact [ENA support](https://www.ebi.ac.uk/ena/browser/support) for guidance. + +- **Coverage calculation from long reads**: Coverage calculation via `coverm` has only been tested and validated with short reads (e.g. Illumina). It has not been tested with long reads (PacBio, Nanopore). Exercise caution when processing data generated from long reads, as issues may occur. + ## Requirements - [Nextflow](https://www.nextflow.io/) `>=25.04.0` From 4452cb8975d5c91f8df5ac1b20e28ce5ed8cd586 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Wed, 8 Jul 2026 14:50:19 +0100 Subject: [PATCH 03/47] refactor limitations block --- README.md | 50 ++++++++++++++++++++++++++++++-------------------- 1 file changed, 30 insertions(+), 20 deletions(-) diff --git a/README.md b/README.md index 7133ca1..1e17529 100644 --- a/README.md +++ b/README.md @@ -31,26 +31,6 @@ Currently, the pipeline supports four submission modes, each routed to a dedicat ![seqsubmit workflow diagram](assets/seqsubmit_schema.png) -## Limitations - -nf-core/seqsubmit does not yet support the following data types and scenarios: - -- **Co-assemblies**: Metagenomic co-assemblies, and MAGs/bins generated from co-assemblies, are not supported. The pipeline currently expects each assembly/MAG/bin to be derived from a single run or an assembly generated from a single run. - -- **Eukaryotic and viral bins/MAGs**: Submitting eukaryotic or viral genomes that require additional metadata generation is not supported. - - For **eukaryotic bins/MAGs**, RNA prediction and quality assessment (completeness/contamination estimation) are not implemented. - - For **viral bins/MAGs**, RNA prediction, quality assessment (completeness/contamination estimation) and taxonomy assignment are not implemented. - -- **Reads in non-FASTQ formats**: Only reads provided as FASTQ (`.fastq.gz`) can be submitted with `--mode reads`. Other formats (e.g. BAM) are not supported as input. - -- **Submission of MAG/bin without submitting the assembly or raw reads**: If a MAG/bin cannot be traced back to an assembly accession or a reads accession — i.e. only a sample accession is available — it cannot currently be submitted, since the pipeline relies on that lineage to build the required metadata. - -- **Submission of assembly without submitting the raw reads**: Similarly, assemblies that have no associated reads accession (`run_accession` column in the sample sheet) are not supported for the same reason. - -- **Single-contig assemblies/MAGs/bins**: Assemblies or MAGs/bins that consist of only one contig cannot be uploaded through this pipeline. ENA classifies these as "chromosomal assemblies", which follow a different submission procedure with different metadata requirements. If you have single-contig assembly/MAG/bin to submit, please contact [ENA support](https://www.ebi.ac.uk/ena/browser/support) for guidance. - -- **Coverage calculation from long reads**: Coverage calculation via `coverm` has only been tested and validated with short reads (e.g. Illumina). It has not been tested with long reads (PacBio, Nanopore). Exercise caution when processing data generated from long reads, as issues may occur. - ## Requirements - [Nextflow](https://www.nextflow.io/) `>=25.04.0` @@ -297,6 +277,36 @@ Key output locations in `--outdir`: For full details, see the [output documentation](https://nf-co.re/seqsubmit/output). +## Limitations + +nf-core/seqsubmit does not yet support the following data types and scenarios, grouped by the mode they affect: + +#### `mags` and `bins` modes (`GENOMESUBMIT`) + +- **Co-assemblies**: MAGs/bins generated from co-assemblies are not supported. The pipeline currently expects each MAG/bin to be derived from an assembly generated from a single run. + +- **Eukaryotic and viral bins/MAGs**: Submitting eukaryotic or viral genomes that require additional metadata generation is not supported. For such genomes tRNA and rRNA prediction, quality assessment (completeness/contamination estimation) and taxonomy assignment are not implemented. If all the required metadata (`completeness`, `contamination`, `stats_generation_software`, `RNA_presence` and `NCBI_lineage`) is already provided in the samplesheet, eukaryotic and viral bins/MAGs can still be submitted normally. + +- **Submission of a MAG/bin without submitting the assembly or raw reads**: If a MAG/bin cannot be traced back to an assembly accession or a reads accession — i.e. only a sample accession is available — it cannot currently be submitted, since the pipeline relies on that lineage to build the required metadata. + +- **Single-contig MAGs/bins**: MAGs/bins that consist of only one contig cannot be uploaded through this pipeline. ENA classifies these as "chromosomal assemblies", which follow a different submission procedure with different metadata requirements. If you have a single-contig MAG/bin to submit, please contact [ENA support](https://www.ebi.ac.uk/ena/browser/support) for guidance. + +- **Coverage calculation from long reads**: `genome_coverage` calculation via `coverm` has only been tested and validated with short reads (e.g. Illumina). It has not been tested with long reads (PacBio, Nanopore). Exercise caution when processing data generated from long reads, as issues may occur. + +#### `metagenomic_assemblies` mode (`ASSEMBLYSUBMIT`) + +- **Co-assemblies**: Metagenomic co-assemblies are not supported. The pipeline currently expects each assembly to be derived from a single run. + +- **Submission of an assembly without submitting the raw reads**: Assemblies that have no associated reads accession (`run_accession` column in the samplesheet) are not supported, since the pipeline relies on that lineage to build the required metadata. + +- **Single-contig assemblies**: Assemblies that consist of only one contig cannot be uploaded through this pipeline. ENA classifies these as "chromosomal assemblies", which follow a different submission procedure with different metadata requirements. If you have a single-contig assembly to submit, please contact [ENA support](https://www.ebi.ac.uk/ena/browser/support) for guidance. + +- **Coverage calculation from long reads**: `coverage` calculation via `coverm` has only been tested and validated with short reads (e.g. Illumina). It has not been tested with long reads (PacBio, Nanopore). Exercise caution when processing data generated from long reads, as issues may occur. + +#### `reads` mode (`READSUBMIT`) + +- **Reads in non-FASTQ formats**: Only reads provided as FASTQ (`.fastq.gz`) can be submitted. Other formats (e.g. BAM) are not supported as input. + ## Credits nf-core/seqsubmit was originally written by [Martin Beracochea](https://github.com/mberacochea), [Ekaterina Sakharova](https://github.com/KateSakharova), [Sofia Ochkalova](https://github.com/ochkalova), [Evangelos Karatzas](https://github.com/vagkaratzas) and [Tim Rozday](https://github.com/timrozday-mgnify). From 6bda9c774448e48ff84b17af3cf00a8154589ff2 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Wed, 8 Jul 2026 15:00:15 +0100 Subject: [PATCH 04/47] add scalability section to README.md --- README.md | 8 ++++++++ 1 file changed, 8 insertions(+) diff --git a/README.md b/README.md index 1e17529..e699631 100644 --- a/README.md +++ b/README.md @@ -277,6 +277,14 @@ Key output locations in `--outdir`: For full details, see the [output documentation](https://nf-co.re/seqsubmit/output). +## Scalability + +Some processes in this pipeline are resource intensive — in particular coverage calculation (`coverm`), quality assessment (`CheckM2`), and taxonomic classification of MAGs/bins (`BAT`). For real life data these steps should be run on an HPC cluster or another compute environment with sufficient CPU, memory, and job scheduling capacity, rather than on a single local machine. + +Depending on the resources available on your machine or cluster, you will likely need to adapt the Nextflow configuration to limit the maximum number of concurrently running jobs and other resource settings (e.g. `max_cpus`, `max_memory`, `max_time`, or `executor.queueSize` in a custom config), so the pipeline doesn't try to claim more resources than are actually available. See the [nf-core configuration documentation](https://nf-co.re/docs/usage/getting_started/configuration) for guidance on writing a custom config for your infrastructure. + +When submitting a large number of records, be aware that ENA recommends limiting submissions to 5000 per day to avoid overfilling their processing queue. If you need to submit more than this, consider splitting your samplesheet and spreading the submission across several days. + ## Limitations nf-core/seqsubmit does not yet support the following data types and scenarios, grouped by the mode they affect: From 9074e16fc2ba4495a48360ade7d84565f82f32f1 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Wed, 8 Jul 2026 15:22:35 +0100 Subject: [PATCH 05/47] refactor contributing guidelines --- docs/CONTRIBUTING.md | 66 +++++--------------------------------------- 1 file changed, 7 insertions(+), 59 deletions(-) diff --git a/docs/CONTRIBUTING.md b/docs/CONTRIBUTING.md index 94bbd9f..e48f6a7 100644 --- a/docs/CONTRIBUTING.md +++ b/docs/CONTRIBUTING.md @@ -184,35 +184,7 @@ If you update images or graphics, follow the nf-core [style guidelines](https:// To ensure consistency and maintainability of the nf-core/seqsubmit pipeline, contributors should follow these guidelines when implementing new features or modifying existing functionality. -1. Understand Workflow Structure - -The pipeline consists of independent workflows, each corresponding to a certain data type: - -- GENOMESUBMIT (mags, bins) -- ASSEMBLYSUBMIT (metagenomic_assemblies) -- READSUBMIT (raw reads) - -When adding new functionality (e.g. a new submission mode such as `isolate_genomes`), ensure that: - -- It integrates cleanly into the appropriate workflow or introduces a well-structured new one. -- Existing workflows remain unaffected unless changes are explicitly required. -- Shared logic is abstracted into reusable modules or subworkflows where possible. - -⸻ - -2. Follow Samplesheet and Schema-Driven Design - -All inputs in seqsubmit are defined via strict samplesheet schemas. - -- Maintain: - - Column order consistency (critical for pipeline execution) - - Backward compatibility where possible -- Avoid duplicating metadata logic—reuse existing parsing and validation patterns. -- New feature should extend or introduce a corresponding schema (`assets/schema_\*.json`) if pipeline inputs are affected. - -⸻ - -3. ENA Submission Requirements +### ENA Submission Requirements Submissions to [ENA](https://www.ebi.ac.uk/ena/browser/home) are governed by a well-defined data model and submission process. These include relationships between entities (e.g. studies, samples, experiments, runs, analyses), required submission steps, and accepted file types and formats. @@ -228,7 +200,7 @@ Contributors should refer directly to the [official ENA documentation](https://e ⸻ -4. Integration with Webin-CLI +### Integration with Webin-CLI All submissions are performed via [Webin-CLI](https://github.com/enasequence/webin-cli). Contributors should rely on official Webin-CLI documentation to understand expected inputs, parameters, and modes of operation. Make sure to always use the latest version of the tool. @@ -242,7 +214,7 @@ All submissions are performed via [Webin-CLI](https://github.com/enasequence/web ⸻ -5. Testing Requirements +### Testing Requirements All new functionality must be covered by tests. @@ -264,36 +236,12 @@ Where applicable: ⸻ -6. Performance and Resource Awareness - -Some data processing steps may be resource-intensive (e.g. database preparation for CAT_pack). When introducing new workflows or features avoid unnecessary recomputation. Add option to reuse cached results or provide prebuilt databases when possible. - -⸻ - -7. Documentation and Usability - -Every contribution should aim to improve usability: - -- Update: - - README.md - - docs/usage.md - - Schema descriptions -- Provide: - - Clear examples (e.g. samplesheets) - - Meaningful parameter descriptions - -The goal is to ensure that users can successfully submit data without needing deep knowledge of ENA submission requirements. - -⸻ - -8. Design Principles +### Performance and Resource Awareness -When contributing, prioritise: +Some data processing steps may be resource-intensive (e.g. database preparation for CAT_pack). -- Reproducibility (consistent results across environments) -- Automation (minimise manual user intervention) -- Clarity (explicit inputs and outputs) -- Extensibility (easy to add new submission types) +- When introducing new workflows or features avoid unnecessary recomputation. +- Add option to reuse cached results or provide prebuilt databases when possible. ⸻ From fe619780915858f190d4b6a4c82a00c68ee19ab4 Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Thu, 9 Jul 2026 14:16:06 +0100 Subject: [PATCH 06/47] start changing --- README.md | 2 +- assets/multiqc_config.yml | 2 +- assets/schema_input_genome.json | 3 +- conf/test_mag_no_quality.conf | 1 + {assets => docs/images}/seqsubmit_schema.png | Bin docs/methods.md | 2 +- docs/usage.md | 64 +++++++++--------- .../local/generate_assembly_manifest/main.nf | 12 +--- modules/local/genome_upload/main.nf | 2 +- nextflow.config | 2 +- subworkflows/local/fasta_validation/main.nf | 4 +- .../utils_nfcore_seqsubmit_pipeline/main.nf | 4 +- workflows/assemblysubmit.nf | 1 - 13 files changed, 45 insertions(+), 54 deletions(-) rename {assets => docs/images}/seqsubmit_schema.png (100%) diff --git a/README.md b/README.md index 40f2f6e..63a854a 100644 --- a/README.md +++ b/README.md @@ -29,7 +29,7 @@ Currently, the pipeline supports four submission modes, each routed to a dedicat - `metagenomic_assemblies` for assembly submission with `ASSEMBLYSUBMIT` workflow - `reads` for raw sequencing reads submission with `READSUBMIT` workflow -![seqsubmit workflow diagram](assets/seqsubmit_schema.png) +![seqsubmit workflow diagram](docs/images/seqsubmit_schema.png) ## Requirements diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index 5df001f..eb16eaf 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -28,7 +28,7 @@ sp: genome_metadata: fn_re: ".*genomes_metadata.tsv$" sample_registration: - fn_re: '.*registered.*\.tsv$' + fn_re: '.*registered\.*\.tsv$' submission_results_genomes: fn_re: 'genomes_accessions\.tsv$' submission_results_assemblies: diff --git a/assets/schema_input_genome.json b/assets/schema_input_genome.json index f851f85..e0aef2b 100644 --- a/assets/schema_input_genome.json +++ b/assets/schema_input_genome.json @@ -91,7 +91,8 @@ }, "co-assembly": { "type": "string", - "description": "Yes if data was co-assembled, No otherwise" + "description": "'Yes' if data was co-assembled, 'No' otherwise", + "pattern": ['Yes'|'No'] }, "broad_environment": { "type": "string", diff --git a/conf/test_mag_no_quality.conf b/conf/test_mag_no_quality.conf index 9d3724a..656bffe 100644 --- a/conf/test_mag_no_quality.conf +++ b/conf/test_mag_no_quality.conf @@ -34,6 +34,7 @@ params { cat_db = null // CheckM2 doesn't allow usage of small test database, so real database will be downloaded during the test run, // which is not ideal but necessary to test the pipeline's behaviour with missing quality values + // Track upstream issue here: https://github.com/chklovski/CheckM2/issues/140 checkm2_db = null } diff --git a/assets/seqsubmit_schema.png b/docs/images/seqsubmit_schema.png similarity index 100% rename from assets/seqsubmit_schema.png rename to docs/images/seqsubmit_schema.png diff --git a/docs/methods.md b/docs/methods.md index 0f15a33..80123ab 100644 --- a/docs/methods.md +++ b/docs/methods.md @@ -98,7 +98,7 @@ Genome coverage values from `coverm genome` output TSV are parsed and merged int ### Taxonomy assignment -If `NCBI_lineage` is already present in the input samlesheet, the value is retained. If it is missing, the workflow runs taxonomy classification using the `CAT_pack` tool. +If `NCBI_lineage` is already present in the input samplesheet, the value is retained. If it is missing, the workflow runs taxonomy classification using the `CAT_pack` tool. Before classification, input FASTA files are normalized to a `.fasta` suffix by `RENAME_FASTA_FOR_CATPACK`. Classification is then run in bin mode (`CAT_pack bins`, followed by `CAT_pack add_names`). diff --git a/docs/usage.md b/docs/usage.md index 7d3b5ef..00274a4 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -51,27 +51,27 @@ mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default, > [!IMPORTANT] > **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. -| Column | Description | -| --------------------------- | ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | A unique identifier for this data entry. Must be globally unique within the input dataset (used as `meta.id` throughout the pipeline). | -| `fasta` | Path to MAG/bin contigs in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. | -| `accession` | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | -| `fastq_1` | Path to the read file in FASTQ format used to generate the source metagenomic assembly. Required if `genome_coverage` is not provided. | -| `fastq_2` | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | -| `assembly_software` | Tool name and version that were used to generate the source metagenomic assembly. | -| `binning_software` | Binning tool, including version, that was used to generate the bins. | -| `binning_parameters` | Arguments that were used during binning. | -| `stats_generation_software` | Tool, including version, that was used to calculate completeness and contamination. | -| `completeness` | Genome completeness value. | -| `contamination` | Genome contamination value. | -| `genome_coverage` | Estimated average sequencing depth across the genome. If the value is missing, it is computed automatically during pipeline execution when reads are provided. | -| `metagenome` | Registered metagenome taxonomic identifier or name that matches an existing ENA taxonomy entry. For more details see https://ena-docs.readthedocs.io/en/latest/faq/taxonomy.html | -| `co-assembly` | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: Yes or No. | -| `broad_environment` | Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428) | -| `local_environment` | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". | +| Column | Description | +| --------------------------- | ---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `sample` | A unique identifier for this data entry. Must be globally unique within the input dataset. | +| `fasta` | Path to MAG/bin contigs in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. | +| `accession` | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | +| `fastq_1` | Path to the read file in FASTQ format used to generate the source metagenomic assembly. Required if `genome_coverage` is not provided. | +| `fastq_2` | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | +| `assembly_software` | Tool name and version that were used to generate the source metagenomic assembly. | +| `binning_software` | Binning tool, including version, that was used to generate the bins. | +| `binning_parameters` | Arguments that were used during binning. | +| `stats_generation_software` | Tool, including version, that was used to calculate completeness and contamination. | +| `completeness` | Genome completeness value. | +| `contamination` | Genome contamination value. | +| `genome_coverage` | Estimated average sequencing depth across the genome. If the value is missing, it is computed automatically during pipeline execution when reads are provided. | +| `metagenome` | Registered metagenome taxonomic identifier or name that matches an existing ENA taxonomy entry. For more details see https://ena-docs.readthedocs.io/en/latest/faq/taxonomy.html | +| `co-assembly` | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: Yes or No. | +| `broad_environment` | Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428) | +| `local_environment` | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". | | `environmental_medium` | Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). | -| `RNA_presence` | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: Yes or No. | -| `NCBI_lineage` | NCBI taxonomy lineage of the genome. Can be composted of either numerical IDs or taxon names separated by ";". | +| `RNA_presence` | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: Yes or No. | +| `NCBI_lineage` | NCBI taxonomy lineage of the genome. Can be composted of either numerical IDs or taxon names separated by ";". | > [!NOTE] > More information about envioronment tags can be found at checklists [ERC000050](https://www.ebi.ac.uk/ena/browser/view/ERC000050) for bins and [ERC000047](https://www.ebi.ac.uk/ena/browser/view/ERC000047) for MAGs under the field names "broad-scale environmental context", "local environmental context", and "environmental medium". @@ -93,16 +93,16 @@ assembly_002,data/assembly_002.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 > [!IMPORTANT] > **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. -| Column | Description | -| ------------------- | ----------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | A unique identifier for this data entry. Must be globally unique within the input dataset (used as `meta.id` throughout the pipeline). | -| `fasta` | Path to assembly contigs in FASTA format compressed with `gzip`. | -| `fastq_1` | Path to the read file in FASTQ format used to generate the metagenomic assembly. Required if `coverage` is not provided. | -| `fastq_2` | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | +| Column | Description | +| ------------------- | ---------------------------------------------------------------------------------------------------------------------------------------------------- | +| `sample` | A unique identifier for this data entry. Must be globally unique within the input dataset. | +| `fasta` | Path to assembly contigs in FASTA format compressed with `gzip`. | +| `fastq_1` | Path to the read file in FASTQ format used to generate the metagenomic assembly. Required if `coverage` is not provided. | +| `fastq_2` | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | | `coverage` | Estimated sequencing depth of the assembly. If this value is missing, it is computed automatically during pipeline execution when reads are provided. | -| `run_accession` | ENA run accession for the reads used to generate the metagenomic assembly. Reads must already be submitted to ENA. | -| `assembler` | Name of the assembler software used to generate the assembly. | -| `assembler_version` | Version of the assembler software used to generate the assembly. | +| `run_accession` | ENA run accession for the reads used to generate the metagenomic assembly. Reads must already be submitted to ENA. | +| `assembler` | Name of the assembler software used to generate the assembly. | +| `assembler_version` | Version of the assembler software used to generate the assembly. | An example file is available at [assets/samplesheet_assembly.csv](../assets/samplesheet_assembly.csv). @@ -246,7 +246,7 @@ Key parameters: | `--webincli_mode` | Webin-CLI mode for ENA interaction: `submit` uploads data, `validate` performs validation only. | | `--upload_tpa` | Mark assemblies as third party assemblies when required. | -Test example for `mags` run with docker: +Example `mags` mode upload to TEST server, run command with docker: ```bash nextflow run nf-core/seqsubmit \ @@ -260,7 +260,7 @@ nextflow run nf-core/seqsubmit \ --outdir results/validate_mags ``` -Test example for `metagenomic_assemblies` run with docker: +Example `metagenomic_assemblies` mode upload to TEST server, run command with docker: ```bash nextflow run nf-core/seqsubmit \ @@ -274,7 +274,7 @@ nextflow run nf-core/seqsubmit \ --outdir results/validate_assemblies ``` -Test example for `reads` run with docker: +Example `reads` mode upload to TEST server, run command with docker: ```bash nextflow run nf-core/seqsubmit \ diff --git a/modules/local/generate_assembly_manifest/main.nf b/modules/local/generate_assembly_manifest/main.nf index 4cc0121..90553c0 100644 --- a/modules/local/generate_assembly_manifest/main.nf +++ b/modules/local/generate_assembly_manifest/main.nf @@ -16,7 +16,7 @@ process GENERATE_ASSEMBLY_MANIFEST { output: tuple val(meta), path("${assembly_study}_upload/*.manifest") , emit: manifest - path "versions.yml", emit: versions + tuple val("${task.process}"), val('assembly_uploader'), eval("assembly_manifest --version | sed 's/assembly_uploader //'"), topic: versions when: task.ext.when == null || task.ext.when @@ -37,11 +37,6 @@ process GENERATE_ASSEMBLY_MANIFEST { ${test_flag} \\ ${is_private_flag} \\ ${args} - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - assembly_uploader: \$(assembly_manifest --version | sed 's/assembly_uploader //') - END_VERSIONS """ stub: @@ -50,10 +45,5 @@ process GENERATE_ASSEMBLY_MANIFEST { """ mkdir ${assembly_study}_upload/ touch ${assembly_study}_upload/test.manifest - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - assembly_uploader: \$(assembly_manifest --version | sed 's/assembly_uploader //') - END_VERSIONS """ } diff --git a/modules/local/genome_upload/main.nf b/modules/local/genome_upload/main.nf index 651b281..095d9ca 100644 --- a/modules/local/genome_upload/main.nf +++ b/modules/local/genome_upload/main.nf @@ -22,7 +22,7 @@ process GENOME_UPLOAD { path "results/{MAG,bin}_upload/ENA_backup.json" , emit: ena_upload_backup_json path "results/{MAG,bin}_upload/genome_samples.xml" , emit: upload_genome_samples path "results/{MAG,bin}_upload/registered_{MAGs,bins}*.tsv", emit: upload_registered_mags - tuple val("${task.process}"), val('genome_uploader'), eval("genome_upload --version 2>&1 | sed 's/genome_uploader //g'"), topic: versions, emit: versions_genome_uploader + tuple val("${task.process}"), val('genome_uploader'), eval("genome_upload --version 2>&1 | sed 's/genome_uploader //g'"), topic: versions when: task.ext.when == null || task.ext.when diff --git a/nextflow.config b/nextflow.config index eaad0b8..2cef81c 100644 --- a/nextflow.config +++ b/nextflow.config @@ -243,7 +243,7 @@ profiles { test { includeConfig 'conf/test.config' } test_full { includeConfig 'conf/test_full.config' } test_mag_complete_metadata { includeConfig 'conf/test_mag_complete_metadata.conf' } - test_mag_no_study_complete_metadata { includeConfig 'conf/test_mag_no_study_complete_metadata.config' } + test_mag_no_study_complete_metadata { includeConfig 'conf/test_mag_no_study_complete_metadata.config' } test_mag_no_coverage_single_reads { includeConfig 'conf/test_mag_no_coverage_single_reads.conf' } test_mag_no_coverage_paired_reads { includeConfig 'conf/test_mag_no_coverage_paired_reads.conf' } test_mag_no_quality { includeConfig 'conf/test_mag_no_quality.conf' } diff --git a/subworkflows/local/fasta_validation/main.nf b/subworkflows/local/fasta_validation/main.nf index 4d2d559..c59df83 100644 --- a/subworkflows/local/fasta_validation/main.nf +++ b/subworkflows/local/fasta_validation/main.nf @@ -8,7 +8,7 @@ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ -include { FALINT } from '../../../modules/nf-core/falint/main' +include { FALINT } from '../../../modules/nf-core/falint/main' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -63,5 +63,5 @@ workflow FASTA_VALIDATION { ) emit: - valid_fastas = valid_fastas + valid_fastas = valid_fastas // [meta, fasta_file] } diff --git a/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf b/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf index 1ac05e9..53fd0dc 100644 --- a/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf @@ -237,12 +237,12 @@ def toolBibliographyText() { def reference_text = [ '
  • Richardson, L., Allen, B., Baldi, G., Beracochea, M., Bileschi, M. L., Burdett, T., ... & Finn, R. D. (2023). MGnify: the microbiome sequence data analysis resource in 2023. Nucleic acids research, 51(D1), D753-D759. doi: 10.1093/nar/gkac1080
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  • Gurbich, T. A., Almeida, A., Beracochea, M., Burdett, T., Burgin, J., Cochrane, G., ... & Finn, R. D. (2023). MGnify genomes: a resource for biome-specific microbial genome catalogues. Journal of molecular biology, 435(14), 168016. doi: 10.1016/j.jmb.2023.168016
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  • webin-cli: David, Y., Alisha, A., Awais, A., Rajkumar, D., Dipayan, G., Muhammad, H., ... & Ugis, S. (2026). The European Nucleotide Archive in 2025. Nucleic Acids Research, 54(D1), D120-D127. doi: 10.1093/nar/gkaf1295
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  • David, Y., Alisha, A., Awais, A., Rajkumar, D., Dipayan, G., Muhammad, H., ... & Ugis, S. (2026). The European Nucleotide Archive in 2025. Nucleic Acids Research, 54(D1), D120-D127. doi: 10.1093/nar/gkaf1295
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  • Edwards, R. (2023). linsalrob/py_fasta_validator: Compressy. doi: 10.5281/zenodo.5002710
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  • Chklovski, A., Parks, D. H., Woodcroft, B. J., & Tyson, G. W. (2023). CheckM2: a rapid, scalable and accurate tool for assessing microbial genome quality using machine learning. Nature Methods, 20(8), 1203-1212. doi: 10.1038/s41592-023-01940-w
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  • Aroney, S. T., Newell, R. J., Nissen, J. N., Camargo, A. P., Tyson, G. W., & Woodcroft, B. J. (2025). CoverM: read alignment statistics for metagenomics. Bioinformatics, 41(4), btaf147. doi: 10.1093/bioinformatics/btaf147
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  • Von Meijenfeldt, F. B., Arkhipova, K., Cambuy, D. D., Coutinho, F. H., & Dutilh, B. E. (2019). Robust taxonomic classification of uncharted microbial sequences and bins with CAT and BAT. Genome biology, 20(1), 217. doi: 10.1038/s41467-024-47155-1
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  • barrnap: Seemann, T. (2014). barrnap: Bacterial ribosomal RNA predictor. barrnap: Bacterial ribosomal RNA predictor. vicbioinformatics.com
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  • Seemann, T. (2014). barrnap: Bacterial ribosomal RNA predictor. barrnap: Bacterial ribosomal RNA predictor. vicbioinformatics.com
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  • Chan, P. P., Lin, B. Y., Mak, A. J., & Lowe, T. M. (2021). tRNAscan-SE 2.0: improved detection and functional classification of transfer RNA genes. Nucleic acids research, 49(16), 9077-9096. doi: 10.1093/nar/gkab688
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  • Ewels, P., Magnusson, M., Lundin, S., & Käller, M. (2016). MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics , 32(19), 3047–3048. doi: 10.1093/bioinformatics/btw354
    • ' ].join(' ').trim() diff --git a/workflows/assemblysubmit.nf b/workflows/assemblysubmit.nf index 84a9dc8..f712b72 100644 --- a/workflows/assemblysubmit.nf +++ b/workflows/assemblysubmit.nf @@ -176,7 +176,6 @@ workflow ASSEMBLYSUBMIT { test_upload, is_private ) - ch_versions = ch_versions.mix(GENERATE_ASSEMBLY_MANIFEST.out.versions.first()) SUBMIT ( assemblies_with_coverage.join(GENERATE_ASSEMBLY_MANIFEST.out.manifest), From 3e9998f4f20f2fd36dd2b6215202071d1e1fba3d Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Thu, 9 Jul 2026 14:30:41 +0100 Subject: [PATCH 07/47] fix --- assets/schema_input_genome.json | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/assets/schema_input_genome.json b/assets/schema_input_genome.json index e0aef2b..26b64ac 100644 --- a/assets/schema_input_genome.json +++ b/assets/schema_input_genome.json @@ -92,7 +92,7 @@ "co-assembly": { "type": "string", "description": "'Yes' if data was co-assembled, 'No' otherwise", - "pattern": ['Yes'|'No'] + "enum": ["Yes", "No"] }, "broad_environment": { "type": "string", From 44c38d9fb33c78ccdaf0964f0fb27f51d0efd013 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Thu, 9 Jul 2026 14:50:50 +0100 Subject: [PATCH 08/47] update CONTRIBUTING.md to clarify testing requirements for ENA submission --- docs/CONTRIBUTING.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/CONTRIBUTING.md b/docs/CONTRIBUTING.md index e48f6a7..7552e65 100644 --- a/docs/CONTRIBUTING.md +++ b/docs/CONTRIBUTING.md @@ -216,7 +216,7 @@ All submissions are performed via [Webin-CLI](https://github.com/enasequence/web ### Testing Requirements -All new functionality must be covered by tests. +All new functionality must be covered by tests. Running nf-tests that perform ENA submission locally requires ENA webin credentials. Contributors may ask for testing credentials on the nf-core Slack [#seqsubmit](https://nfcore.slack.com/channels/seqsubmit) channel. At minimum, include: From 67d436af06f56a0af45c15fa7ac1885bbc2628a1 Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Thu, 9 Jul 2026 15:02:32 +0100 Subject: [PATCH 09/47] add some links --- assets/schema_input_genome.json | 23 ++++++++++++++--------- 1 file changed, 14 insertions(+), 9 deletions(-) diff --git a/assets/schema_input_genome.json b/assets/schema_input_genome.json index 26b64ac..65472dc 100644 --- a/assets/schema_input_genome.json +++ b/assets/schema_input_genome.json @@ -75,11 +75,15 @@ }, "completeness": { "type": "number", - "description": "MAG/bin completeness value" + "description": "MAG/bin completeness score: the ratio of observed single-copy marker genes to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined", + "exclusiveMinimum": 0, + "exclusiveMaximum": 100 }, "contamination": { "type": "number", - "description": "MAG/bin contamination value" + "description": "MAG/bin contamination score: the ratio of observed single-copy marker genes in ≥2 copies to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined", + "exclusiveMinimum": 0, + "exclusiveMaximum": 100 }, "genome_coverage": { "type": "number", @@ -87,32 +91,33 @@ }, "metagenome": { "type": "string", - "description": "ENA accepted metagenome name" + "description": "ENA accepted metagenome name. It needs to be listed in the taxonomy tree https://www.ebi.ac.uk/ena/browser/view/408169?show=tax-tree (you might need to press \"Tax tree - Show\" in the right most section of the page). Full list can also be found https://github.com/EBI-Metagenomics/genome_uploader/blob/main/genomeuploader/constants.py#L22" }, "co-assembly": { "type": "string", - "description": "'Yes' if data was co-assembled, 'No' otherwise", + "description": "'Yes' if data was co-assembled, 'No' otherwise. Options: 'Yes' or 'No'", "enum": ["Yes", "No"] }, "broad_environment": { "type": "string", - "description": "broad ecological context of a sample" + "description": "Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428). Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS" }, "local_environment": { "type": "string", - "description": "local ecological context" + "description": "Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for \"broad-scale environmental context\". Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS" }, "environmental_medium": { "type": "string", - "description": "material displaced by the sample" + "description": "Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS" }, "RNA_presence": { "type": "string", - "description": "Yes/No if at least 18 tRNAs and 5S, 16S, 23S rRNA genes detected" + "description": "Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: 'Yes' or 'No'.", + "enum": ["Yes", "No"] }, "NCBI_lineage": { "type": "string", - "description": "full NCBI lineage - format: x;y;z" + "description": "full NCBI lineage - format: x;y;z. For example, the lineage for E. coli can be: 'Bacteria;Pseudomonadati;Pseudomonadota;Gammaproteobacteria;Enterobacterales;Enterobacteriaceae;Escherichia' or '2;1224;1236;91347;543;561;562'" } }, "required": [ From b0343f445ce70c0933beb76eb6ba72d69b5b4734 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Thu, 9 Jul 2026 15:07:00 +0100 Subject: [PATCH 10/47] update README.md to reference Methods documentation for metadata statistics and coverage calculation --- README.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/README.md b/README.md index e699631..07b3f72 100644 --- a/README.md +++ b/README.md @@ -80,7 +80,7 @@ Additional supported columns: - `RNA_presence` - `NCBI_lineage` -If `genome_coverage`, `stats_generation_software`, `completeness`, `contamination`, `RNA_presence`, or `NCBI_lineage` are missing, the workflow can calculate or infer them when the required inputs are available. +If `genome_coverage`, `stats_generation_software`, `completeness`, `contamination`, `RNA_presence`, or `NCBI_lineage` are missing, the workflow can calculate or infer them when the required inputs are available. See the [Methods documentation](docs/methods.md) for more information on how metadata statistics are obtained. Those fields are metadata required for the [genome_uploader](https://github.com/EBI-Metagenomics/genome_uploader) package. @@ -111,7 +111,7 @@ At least one of the following must be provided per row: - reads (`fastq_1`, optional `fastq_2` for paired-end) - `coverage` -If `coverage` is missing and reads are provided, the workflow calculates average coverage with `coverm`. +If `coverage` is missing and reads are provided, the workflow calculates average coverage with `coverm`. See the [Methods documentation](docs/methods.md) for more information on how coverage is calculated. Example `samplesheet_assembly.csv`: From f7f99b32df13817b972d0a73b708dae848504b2b Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Thu, 9 Jul 2026 15:07:23 +0100 Subject: [PATCH 11/47] refactor methods documentation to clarify RNA presence detection and update metadata schema --- docs/methods.md | 50 ++++++++----------- .../local/create_genome_metadata_tsv/meta.yml | 2 +- nextflow_schema.json | 8 +-- 3 files changed, 25 insertions(+), 35 deletions(-) diff --git a/docs/methods.md b/docs/methods.md index 0f15a33..acbf12f 100644 --- a/docs/methods.md +++ b/docs/methods.md @@ -1,15 +1,8 @@ # nf-core/seqsubmit: Methods -## Overview +This page provides a detailed overview of the methods and procedures that are executed throughout the nf-core/seqsubmit pipeline. -`nf-core/seqsubmit` currently contains two workflow implementations: - -- `GENOMESUBMIT` for `--mode mags` and `--mode bins` -- `ASSEMBLYSUBMIT` for `--mode metagenomic_assemblies` - -This page documents the methods that are currently implemented in the pipeline. - -## `GENOMESUBMIT` methods +## `GENOMESUBMIT` ### Overview @@ -17,22 +10,24 @@ The `GENOMESUBMIT` workflow: 1. Reads the samplesheet and associated genome FASTA files. 2. Validates genome FASTA files. -3. Reuses provided or calculates missing values for RNA genes presence, coverage, taxonomy, and genome quality metrics. +3. Reuses provided or calculates missing values for tRNA and rRNA genes presence, coverage, taxonomy, and genome quality metrics. 4. Collects genome metadata into the tabular format required by `genome_uploader`. 5. Generates submission manifests for ENA. 6. Performs submission to ENA. ### Genome FASTA validation -Genome FASTA files are validated with the `FASTAVALIDATOR` module before downstream processing. Each file is checked for FASTA format validity and contig count. A genome must contain at least two contigs to pass validation, which is an ENA requirement for contig-level submissions. +Genome FASTA files are validated with the `fa-lint` before downstream processing. Each file is checked for FASTA format validity and contig count. A genome must contain at least two contigs to pass validation, which is an ENA requirement for contig-level submissions. Only FASTA files that pass validation are retained for downstream processing and submission. -### RNA presence detection +### tRNA and rRNA genes detection -The workflow only runs internal RNA detection for entries where the `RNA_presence` column is empty. If a value is already supplied in the samplesheet, that value is passed through unchanged. +The workflow only runs tRNA and rRNA genes detection for entries where the `RNA_presence` column is empty. If a value is already supplied in the samplesheet, that value is passed through unchanged. -RNA detection is implemented through the `RNA_DETECTION` subworkflow and combines: +The resulting `RNA_presence` value is a decisive factor affecting MISAG/MIMAG assembly quality classification of a genome. + +tRNA and rRNA genes detection is implemented through the `RNA_DETECTION` subworkflow and combines: - `barrnap` for rRNA prediction - `tRNAscan-SE` for tRNA prediction @@ -42,11 +37,7 @@ RNA detection is implemented through the `RNA_DETECTION` subworkflow and combine `barrnap` is run in bacterial mode (`"bac"`). -The custom parser then scans the GFF output and keeps only the following ribosomal RNA subunits: - -- `16S_rRNA` -- `23S_rRNA` -- `5S_rRNA` +The custom parser then scans the GFF output and keeps only the following rRNA subunits: `16S_rRNA`, `23S_rRNA`, `5S_rRNA`. For each detected feature, the recovered length is calculated as: @@ -66,7 +57,7 @@ $$ If multiple hits are found for the same subunit, the workflow keeps the best recovered percentage for that subunit. -A subunit is considered present when its best recovered percentage is greater than or equal to `--rrna_limit`. The current default is `80`. +A subunit is considered present when its best recovered percentage is greater than or equal to `--rrna_limit`. The current default is `80`. This threshold is arbitrary, as no strict consensus length is defined by ENA or MISAG/MIMAG for considering an rRNA gene present. #### tRNA detection @@ -79,16 +70,16 @@ The parser sums the counts assigned to the 20 standard amino-acid isotypes: - `Leu`, `Lys`, `Met`, `Phe`, `Pro` - `Ser`, `Thr`, `Trp`, `Tyr`, `Val` -The total number of predicted tRNAs is compared against `params.trna_limit`. The current default is `18`. +The total number of predicted tRNAs is compared against `params.trna_limit`. The current default is `18`. This value is defined by the MISAG/MIMAG standard and must not be modified. -#### Final RNA presence decision +#### Final `RNA_presence` decision The final decision stored in `RNA_presence` is: - `Yes` when at least 18 tRNAs are detected and all three required rRNA subunits pass the recovery threshold - `No` otherwise -The result is written as a two-column TSV file containing the genome identifier and the final `Yes`/`No` decision, and this value is then merged back into the submission metadata. +The final `Yes`/`No` decision is then merged back into the submission metadata. ### Genome coverage calculation @@ -98,7 +89,7 @@ Genome coverage values from `coverm genome` output TSV are parsed and merged int ### Taxonomy assignment -If `NCBI_lineage` is already present in the input samlesheet, the value is retained. If it is missing, the workflow runs taxonomy classification using the `CAT_pack` tool. +If `NCBI_lineage` is already present in the input samplesheet, the value is retained. If it is missing, the workflow runs taxonomy classification using the `CAT_pack` tool. Before classification, input FASTA files are normalized to a `.fasta` suffix by `RENAME_FASTA_FOR_CATPACK`. Classification is then run in bin mode (`CAT_pack bins`, followed by `CAT_pack add_names`). @@ -112,7 +103,7 @@ The workflow checks three samplesheet fields: `completeness`, `contamination`, a For records that run `CheckM2`, completeness and contamination are extracted from the generated quality report (`quality_report.tsv`) and used `CheckM2` version is recorded as `stats_generation_software`. -## `ASSEMBLYSUBMIT` methods +## `ASSEMBLYSUBMIT` ### Overview @@ -120,14 +111,13 @@ The `ASSEMBLYSUBMIT` workflow: 1. Reads the samplesheet and associated assembly FASTA files. 2. Validates assembly FASTA files. -3. Reuses `coverage` values supplied in the samplesheet when they are already present. -4. Calculates `coverage` internally for entries where this field is missing and reads are available. -5. Builds the metadata table used for manifest generation. -6. Generates assembly manifests and submits the assemblies to ENA. +3. Reuses provided or calculates missing `coverage` values. +4. Builds the metadata table used for manifest generation. +5. Generates assembly manifests and submits the assemblies to ENA. ### Assembly FASTA validation -Assembly FASTA files are validated with `FASTAVALIDATOR` before downstream processing. Each file is checked for FASTA format validity and contig count. A genome must contain at least two contigs to pass validation, which is an ENA requirement for contig-level submissions. +Assembly FASTA files are validated with `fa-lint` before downstream processing. Each file is checked for FASTA format validity and contig count. A genome must contain at least two contigs to pass validation, which is an ENA requirement for contig-level submissions. Only assemblies with successful validation are forwarded to coverage estimation, metadata/manifest generation and submission. diff --git a/modules/local/create_genome_metadata_tsv/meta.yml b/modules/local/create_genome_metadata_tsv/meta.yml index 74752ad..870b889 100644 --- a/modules/local/create_genome_metadata_tsv/meta.yml +++ b/modules/local/create_genome_metadata_tsv/meta.yml @@ -35,7 +35,7 @@ input: - broad_environment: Broad environmental context - local_environment: Local environmental context - environmental_medium: Environmental medium description - - RNA_presence: RNA presence status ("Yes" or other) + - RNA_presence: status of tRNA and rRNA genes presence ("Yes" or other) - NCBI_lineage: NCBI taxonomic lineage - fasta: type: file diff --git a/nextflow_schema.json b/nextflow_schema.json index 59c6dc0..b7d7da5 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -43,11 +43,11 @@ } }, "rna_detection_options": { - "title": "RNA detection options", + "title": "rRNA and tRNA detection options", "type": "object", "fa_icon": "fas fa-university", - "description": "Parameters used to make a decision about presence/absence of RNA in data", - "help_text": "ENA requires a boolean value (True/False) defining RNA. Decision of presence/absence of RNA is based on number of rRNA and tRNA. If both values are bigger than their corresponding limits - RNA is present (True), otherwise - not (False).", + "description": "Parameters affecting a decision about presence/absence of tRNA and rRNA genes (RNA_presence) in a genome", + "help_text": "The value of RNA_presence is based on the number of rRNA and tRNA. If at least 18 tRNAs are detected and 16S, 23S, and 5S rRNA subunits pass the recovery threshold - RNA_presence is True, otherwise - False. The resulting value is a decisive factor affecting MISAG/MIMAG assembly quality classification of a genome.", "properties": { "rrna_limit": { "type": "number", @@ -57,7 +57,7 @@ }, "trna_limit": { "type": "number", - "description": "Minimum number of tRNA.", + "description": "Minimum number of tRNA genes detected to count tRNA as present. This value is defined by the MISAG/MIMAG standard and must not be modified.", "default": 18, "fa_icon": "fas fa-users-cog" } From 36567f412f06f80241e09396939c27729ed09fe0 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Thu, 9 Jul 2026 15:09:19 +0100 Subject: [PATCH 12/47] add SVG pipeline metromap --- assets/seqsubmit_schema.svg | 114 ++++++++++++++++++++++++++++++++++++ 1 file changed, 114 insertions(+) create mode 100644 assets/seqsubmit_schema.svg diff --git a/assets/seqsubmit_schema.svg b/assets/seqsubmit_schema.svg new file mode 100644 index 0000000..2338fce --- /dev/null +++ b/assets/seqsubmit_schema.svg @@ -0,0 +1,114 @@ + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + From c57e86589021709490079d438a7a2e6c03c8872e Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Fri, 10 Jul 2026 11:19:09 +0100 Subject: [PATCH 13/47] update README.md Limitations to include issue tracking --- README.md | 10 +++++----- 1 file changed, 5 insertions(+), 5 deletions(-) diff --git a/README.md b/README.md index 07b3f72..db00849 100644 --- a/README.md +++ b/README.md @@ -291,11 +291,11 @@ nf-core/seqsubmit does not yet support the following data types and scenarios, g #### `mags` and `bins` modes (`GENOMESUBMIT`) -- **Co-assemblies**: MAGs/bins generated from co-assemblies are not supported. The pipeline currently expects each MAG/bin to be derived from an assembly generated from a single run. +- **Co-assemblies**: MAGs/bins generated from co-assemblies are not supported. The pipeline currently expects each MAG/bin to be derived from an assembly generated from a single run. Tracked in [issue #61](https://github.com/nf-core/seqsubmit/issues/61). -- **Eukaryotic and viral bins/MAGs**: Submitting eukaryotic or viral genomes that require additional metadata generation is not supported. For such genomes tRNA and rRNA prediction, quality assessment (completeness/contamination estimation) and taxonomy assignment are not implemented. If all the required metadata (`completeness`, `contamination`, `stats_generation_software`, `RNA_presence` and `NCBI_lineage`) is already provided in the samplesheet, eukaryotic and viral bins/MAGs can still be submitted normally. +- **Eukaryotic and viral bins/MAGs**: Submitting eukaryotic or viral genomes that require additional metadata generation is not supported. For such genomes tRNA and rRNA prediction, quality assessment (completeness/contamination estimation) and taxonomy assignment are not implemented. If all the required metadata (`completeness`, `contamination`, `stats_generation_software`, `RNA_presence` and `NCBI_lineage`) is already provided in the samplesheet, eukaryotic and viral bins/MAGs can still be submitted normally. Tracked in [issue #40](https://github.com/nf-core/seqsubmit/issues/40) (eukaryotic) and [issue #63](https://github.com/nf-core/seqsubmit/issues/63) (viral). -- **Submission of a MAG/bin without submitting the assembly or raw reads**: If a MAG/bin cannot be traced back to an assembly accession or a reads accession — i.e. only a sample accession is available — it cannot currently be submitted, since the pipeline relies on that lineage to build the required metadata. +- **Submission of a MAG/bin without submitting the assembly or raw reads**: If a MAG/bin cannot be traced back to an assembly accession or a reads accession — i.e. only a sample accession is available — it cannot currently be submitted, since the pipeline relies on that lineage to build the required metadata. Tracked in [issue #87](https://github.com/nf-core/seqsubmit/issues/87). - **Single-contig MAGs/bins**: MAGs/bins that consist of only one contig cannot be uploaded through this pipeline. ENA classifies these as "chromosomal assemblies", which follow a different submission procedure with different metadata requirements. If you have a single-contig MAG/bin to submit, please contact [ENA support](https://www.ebi.ac.uk/ena/browser/support) for guidance. @@ -303,9 +303,9 @@ nf-core/seqsubmit does not yet support the following data types and scenarios, g #### `metagenomic_assemblies` mode (`ASSEMBLYSUBMIT`) -- **Co-assemblies**: Metagenomic co-assemblies are not supported. The pipeline currently expects each assembly to be derived from a single run. +- **Co-assemblies**: Metagenomic co-assemblies are not supported. The pipeline currently expects each assembly to be derived from a single run. Tracked in [issue #61](https://github.com/nf-core/seqsubmit/issues/61), addressed in [PR #66](https://github.com/nf-core/seqsubmit/pull/66). -- **Submission of an assembly without submitting the raw reads**: Assemblies that have no associated reads accession (`run_accession` column in the samplesheet) are not supported, since the pipeline relies on that lineage to build the required metadata. +- **Submission of an assembly without submitting the raw reads**: Assemblies that have no associated reads accession (`run_accession` column in the samplesheet) are not supported, since the pipeline relies on that lineage to build the required metadata. Tracked in [issue #29](https://github.com/nf-core/seqsubmit/issues/29). - **Single-contig assemblies**: Assemblies that consist of only one contig cannot be uploaded through this pipeline. ENA classifies these as "chromosomal assemblies", which follow a different submission procedure with different metadata requirements. If you have a single-contig assembly to submit, please contact [ENA support](https://www.ebi.ac.uk/ena/browser/support) for guidance. From f759e9c2f76fa86d860be68f70867502ef004b66 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Fri, 10 Jul 2026 11:36:48 +0100 Subject: [PATCH 14/47] update README.md to add a Table of Contents --- README.md | 20 +++++++++++++++++++- 1 file changed, 19 insertions(+), 1 deletion(-) diff --git a/README.md b/README.md index db00849..9fb879a 100644 --- a/README.md +++ b/README.md @@ -19,6 +19,24 @@ [![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23seqsubmit-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/seqsubmit)[![Follow on Bluesky](https://img.shields.io/badge/bluesky-%40nf__core-1185fe?labelColor=000000&logo=bluesky)](https://bsky.app/profile/nf-co.re)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core) +## Table of Contents + +- [Introduction](#introduction) +- [Requirements](#requirements) +- [Input samplesheets](#input-samplesheets) +- [Usage](#usage) + - [Submission study](#submission-study) + - [Data privacy](#data-privacy) + - [Database setup (`CheckM2` and `CAT_pack`)](#database-setup-checkm2-and-cat_pack) + - [Required parameters:](#required-parameters) + - [Optional parameters:](#optional-parameters) +- [Pipeline output](#pipeline-output) +- [Scalability](#scalability) +- [Limitations](#limitations) +- [Credits](#credits) +- [Contributions and Support](#contributions-and-support) +- [Citations](#citations) + ## Introduction **nf-core/seqsubmit** is a Nextflow pipeline for submitting sequence data to [ENA](https://www.ebi.ac.uk/ena/browser/home). @@ -205,7 +223,7 @@ The `mags`/`bins` workflow requires databases for completeness/contamination est | `--upload_tpa` | Flag to control the type of assembly study (third party assembly or not). Default: false | | `--test_upload` | Upload to TEST ENA server instead of LIVE. Default: true | | `--webincli_mode` | Choose Webin-CLI mode: `submit` or `validate`. Default: `submit` | -| `--private` | Use that flag if you are referring to private data in ENA | +| `--is_private` | Use that flag if you are referring private data accessions in your submission | | `--release_date` | Use that flag if you want to keep your data private after submission until particular date | General command template: From a7f6dd63fd0e884ccbd89a8bc7cc0e00bb49d372 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Fri, 10 Jul 2026 11:59:12 +0100 Subject: [PATCH 15/47] Move Data privacy section to usage.md --- README.md | 15 +-------------- docs/usage.md | 17 +++++++++++++++++ 2 files changed, 18 insertions(+), 14 deletions(-) diff --git a/README.md b/README.md index 9fb879a..1a8fd84 100644 --- a/README.md +++ b/README.md @@ -187,20 +187,7 @@ See the [usage documentation](docs/usage.md#submission-study) for more details. ### Data privacy -You can reference private ENA data if you have access to it via your Webin account (it was submitted previously under your credentials). To use private data accessions in your submission metadata, specify the `--is_private` flag. The pipeline will then use your provided Webin credentials to fetch the required metadata. - -You can also control the privacy of your submitted data: - -- If you provide an existing study via `--submission_study`, your submission will inherit the same privacy status as that study. -- If no study is provided, the pipeline will register a new one for you. To keep this new study private, you must specify a release date using `--release_date YYYY-MM-DD` (up to 2 years from the current date). - -| Example | Source Data | Submission Visibility | Required Arguments | Result | -| --------------------------------------------------------------------------------------------------------------------------------------------------------------------- | ----------- | --------------------- | ------------------------------------------------------ | ----------------------------------------------------- | -| Assemblies generated from public reads. Assemblies should be public immediately after submission | public | public | – | SUCCESS | -| Assemblies generated from public reads. Assemblies should remain private for 1 year after submission | public | private | `--release_date YYYY-MM-DD` (+1 year) | SUCCESS | -| Bins generated from private reads and assemblies. Bins should be public after submission. User **has access** to the reads and assemblies via Webin account | private | public | `--is_private` | SUCCESS | -| Bins generated from private reads and assemblies. Bins should remain private for 2 years after submission. User **has access** via Webin account | private | private | `--is_private`, `--release_date YYYY-MM-DD` (+2 years) | SUCCESS | -| MAGs generated from private reads and assemblies. MAGs should be public after submission. User **does not have access** to the reads and assemblies via Webin account | private | public | – | ERROR (submission can only reference accessible data) | +The pipeline supports private submissions. See the [usage documentation](docs/usage.md#data-privacy) for more details. ### Database setup (`CheckM2` and `CAT_pack`) diff --git a/docs/usage.md b/docs/usage.md index 7d3b5ef..54177c5 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -198,6 +198,23 @@ study-soil-2026 Soil microbiome study Survey of soil microbiota | `existing_study_type` | No | ENA study type (e.g. `Metagenomics`, `Other`). | | `new_study_type` | No | Custom study type. Only used when `existing_study_type` is set to `Other`. | +## Data privacy + +You can reference private ENA data if you have access to it via your Webin account (it was submitted previously under your credentials). To use private data accessions in your submission metadata, specify the `--is_private` flag. The pipeline will then use your provided Webin credentials to fetch the required metadata. + +You can also control the privacy of your submitted data: + +- If you provide an existing study via `--submission_study`, your submission will inherit the same privacy status as that study. +- If no study is provided, the pipeline will register a new one for you. To keep this new study private, you must specify a release date using `--release_date YYYY-MM-DD` (up to 2 years from the current date). + +| Example | Source Data | Submission Visibility | Required Arguments | Result | +| --------------------------------------------------------------------------------------------------------------------------------------------------------------------- | ----------- | --------------------- | ------------------------------------------------------ | ----------------------------------------------------- | +| Assemblies generated from public reads. Assemblies should be public immediately after submission | public | public | – | SUCCESS | +| Assemblies generated from public reads. Assemblies should remain private for 1 year after submission | public | private | `--release_date YYYY-MM-DD` (+1 year) | SUCCESS | +| Bins generated from private reads and assemblies. Bins should be public after submission. User **has access** to the reads and assemblies via Webin account | private | public | `--is_private` | SUCCESS | +| Bins generated from private reads and assemblies. Bins should remain private for 2 years after submission. User **has access** via Webin account | private | private | `--is_private`, `--release_date YYYY-MM-DD` (+2 years) | SUCCESS | +| MAGs generated from private reads and assemblies. MAGs should be public after submission. User **does not have access** to the reads and assemblies via Webin account | private | public | – | ERROR (submission can only reference accessible data) | + ## Database preparation (`mags` / `bins`) The `GENOMESUBMIT` workflow uses `CheckM2` and `CAT_pack` that require specialized databases for completeness/contamination assessment and taxonomy assignment. From 81e64bdb908cdcad2bb2f98add04ce0db1aaa8c1 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Fri, 10 Jul 2026 12:11:00 +0100 Subject: [PATCH 16/47] use one format for tables with column descriptions in usage.md --- docs/usage.md | 90 +++++++++++++++++++++++++-------------------------- 1 file changed, 45 insertions(+), 45 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index 54177c5..bc30044 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -51,27 +51,27 @@ mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default, > [!IMPORTANT] > **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. -| Column | Description | -| --------------------------- | ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | A unique identifier for this data entry. Must be globally unique within the input dataset (used as `meta.id` throughout the pipeline). | -| `fasta` | Path to MAG/bin contigs in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. | -| `accession` | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | -| `fastq_1` | Path to the read file in FASTQ format used to generate the source metagenomic assembly. Required if `genome_coverage` is not provided. | -| `fastq_2` | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | -| `assembly_software` | Tool name and version that were used to generate the source metagenomic assembly. | -| `binning_software` | Binning tool, including version, that was used to generate the bins. | -| `binning_parameters` | Arguments that were used during binning. | -| `stats_generation_software` | Tool, including version, that was used to calculate completeness and contamination. | -| `completeness` | Genome completeness value. | -| `contamination` | Genome contamination value. | -| `genome_coverage` | Estimated average sequencing depth across the genome. If the value is missing, it is computed automatically during pipeline execution when reads are provided. | -| `metagenome` | Registered metagenome taxonomic identifier or name that matches an existing ENA taxonomy entry. For more details see https://ena-docs.readthedocs.io/en/latest/faq/taxonomy.html | -| `co-assembly` | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: Yes or No. | -| `broad_environment` | Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428) | -| `local_environment` | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". | -| `environmental_medium` | Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). | -| `RNA_presence` | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: Yes or No. | -| `NCBI_lineage` | NCBI taxonomy lineage of the genome. Can be composted of either numerical IDs or taxon names separated by ";". | +| Column | Required | Description | +| --------------------------- | ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `sample` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset (used as `meta.id` throughout the pipeline). | +| `fasta` | Yes | Path to MAG/bin contigs in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. | +| `accession` | Yes | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | +| `fastq_1` | Conditional | Path to the read file in FASTQ format used to generate the source metagenomic assembly. Required if `genome_coverage` is not provided. | +| `fastq_2` | No | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | +| `assembly_software` | Yes | Tool name and version that were used to generate the source metagenomic assembly. | +| `binning_software` | Yes | Binning tool, including version, that was used to generate the bins. | +| `binning_parameters` | Yes | Arguments that were used during binning. | +| `stats_generation_software` | No | Tool, including version, that was used to calculate completeness and contamination. | +| `completeness` | No | Genome completeness value. | +| `contamination` | No | Genome contamination value. | +| `genome_coverage` | Conditional | Estimated average sequencing depth across the genome. If the value is missing, it is computed automatically during pipeline execution when reads are provided. | +| `metagenome` | Yes | Registered metagenome taxonomic identifier or name that matches an existing ENA taxonomy entry. For more details see https://ena-docs.readthedocs.io/en/latest/faq/taxonomy.html | +| `co-assembly` | Yes | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: Yes or No. | +| `broad_environment` | Yes | Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428) | +| `local_environment` | Yes | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". | +| `environmental_medium` | Yes | Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). | +| `RNA_presence` | No | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: Yes or No. | +| `NCBI_lineage` | No | NCBI taxonomy lineage of the genome. Can be composted of either numerical IDs or taxon names separated by ";". | > [!NOTE] > More information about envioronment tags can be found at checklists [ERC000050](https://www.ebi.ac.uk/ena/browser/view/ERC000050) for bins and [ERC000047](https://www.ebi.ac.uk/ena/browser/view/ERC000047) for MAGs under the field names "broad-scale environmental context", "local environmental context", and "environmental medium". @@ -93,16 +93,16 @@ assembly_002,data/assembly_002.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 > [!IMPORTANT] > **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. -| Column | Description | -| ------------------- | ----------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | A unique identifier for this data entry. Must be globally unique within the input dataset (used as `meta.id` throughout the pipeline). | -| `fasta` | Path to assembly contigs in FASTA format compressed with `gzip`. | -| `fastq_1` | Path to the read file in FASTQ format used to generate the metagenomic assembly. Required if `coverage` is not provided. | -| `fastq_2` | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | -| `coverage` | Estimated sequencing depth of the assembly. If this value is missing, it is computed automatically during pipeline execution when reads are provided. | -| `run_accession` | ENA run accession for the reads used to generate the metagenomic assembly. Reads must already be submitted to ENA. | -| `assembler` | Name of the assembler software used to generate the assembly. | -| `assembler_version` | Version of the assembler software used to generate the assembly. | +| Column | Required | Description | +| ------------------- | ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------- | +| `sample` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset (used as `meta.id` throughout the pipeline). | +| `fasta` | Yes | Path to assembly contigs in FASTA format compressed with `gzip`. | +| `fastq_1` | Conditional | Path to the read file in FASTQ format used to generate the metagenomic assembly. Required if `coverage` is not provided. | +| `fastq_2` | No | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | +| `coverage` | Conditional | Estimated sequencing depth of the assembly. If this value is missing, it is computed automatically during pipeline execution when reads are provided. | +| `run_accession` | Yes | ENA run accession for the reads used to generate the metagenomic assembly. Reads must already be submitted to ENA. | +| `assembler` | Yes | Name of the assembler software used to generate the assembly. | +| `assembler_version` | Yes | Version of the assembler software used to generate the assembly. | An example file is available at [assets/samplesheet_assembly.csv](../assets/samplesheet_assembly.csv). @@ -121,20 +121,20 @@ pacbio_run_001,SAMEA7654321,data/pacbio_reads.fastq.gz,,PACBIO_SMRT,PacBio Seque > [!IMPORTANT] > **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. -| Column | Type | Required | Description | -| ------------------- | --------- | -------- | --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | str | Yes | Unique identifier of this particular data entry. Used as an experiment name. | -| `sample_accession` | str | Yes | ENA sample accession (starting with SAMEA) of the sample used to generate raw reads. | -| `fastq_1` | file path | Yes | Path to forward reads in FASTQ format (optionally gzipped). | -| `fastq_2` | file path | No | Path to reverse reads for paired-end data. Leave empty for single-end reads. | -| `platform` | str | Yes | Sequencing platform. Supported values: `ILLUMINA`, `PACBIO_SMRT`, `OXFORD_NANOPORE`, `ION_TORRENT`, `CAPILLARY`, `DNBSEQ`, `ELEMENT`, `GENAPSYS`, `GENEMIND`, `HELICOS`, `LS454`, `BGISEQ`, `ULTIMA`, `VELA_DIAGNOSTICS`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | -| `instrument` | str | Yes | Sequencer model, e.g. "Illumina HiSeq 2000", "PacBio Sequel", "MinION". | -| `library_source` | str | Yes | Library source type. Options: `GENOMIC`, `METAGENOMIC`, `TRANSCRIPTOMIC`, `METAGENOMIC SINGLE CELL`, `TRANSCRIPTOMIC SINGLE CELL`, `SYNTHETIC`, `VIRAL RNA`, `OTHER`. | -| `library_selection` | str | Yes | Library selection method. Options: `RANDOM`, `PCR`, `RANDOM PCR`, `RT-PCR`, `MF`, `cDNA`, `cDNA_randomPriming`, `cDNA_oligo_dT`, `PolyA`, `Inverse rRNA`, `ChIP`, `MNase`, `DNase`, `Hybrid Selection`, etc. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | -| `library_strategy` | str | Yes | Library strategy. Options: `WGS`, `WGA`, `WXS`, `RNA-Seq`, `miRNA-Seq`, `ncRNA-Seq`, `EST`, `Hi-C`, `ATAC-seq`, `WCS`, `RAD-Seq`, `CLONE`, `AMPLICON`, `POOLCLONE`, `etc`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | -| `insert_size` | number | No | Fragment/insert size for paired-end reads (e.g., 500 for 500 bp inserts). Leave empty if not applicable. | -| `library_name` | str | No | Descriptive library name (optional). | -| `description` | str | No | Free-text description of the experiment (optional). | +| Column | Required | Description | +| ------------------- | -------- | --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `sample` | Yes | Unique identifier of this particular data entry. Used as an experiment name. | +| `sample_accession` | Yes | ENA sample accession (starting with SAMEA) of the sample used to generate raw reads. | +| `fastq_1` | Yes | Path to forward reads in FASTQ format (optionally gzipped). | +| `fastq_2` | No | Path to reverse reads for paired-end data. Leave empty for single-end reads. | +| `platform` | Yes | Sequencing platform. Supported values: `ILLUMINA`, `PACBIO_SMRT`, `OXFORD_NANOPORE`, `ION_TORRENT`, `CAPILLARY`, `DNBSEQ`, `ELEMENT`, `GENAPSYS`, `GENEMIND`, `HELICOS`, `LS454`, `BGISEQ`, `ULTIMA`, `VELA_DIAGNOSTICS`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | +| `instrument` | Yes | Sequencer model, e.g. "Illumina HiSeq 2000", "PacBio Sequel", "MinION". | +| `library_source` | Yes | Library source type. Options: `GENOMIC`, `METAGENOMIC`, `TRANSCRIPTOMIC`, `METAGENOMIC SINGLE CELL`, `TRANSCRIPTOMIC SINGLE CELL`, `SYNTHETIC`, `VIRAL RNA`, `OTHER`. | +| `library_selection` | Yes | Library selection method. Options: `RANDOM`, `PCR`, `RANDOM PCR`, `RT-PCR`, `MF`, `cDNA`, `cDNA_randomPriming`, `cDNA_oligo_dT`, `PolyA`, `Inverse rRNA`, `ChIP`, `MNase`, `DNase`, `Hybrid Selection`, etc. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | +| `library_strategy` | Yes | Library strategy. Options: `WGS`, `WGA`, `WXS`, `RNA-Seq`, `miRNA-Seq`, `ncRNA-Seq`, `EST`, `Hi-C`, `ATAC-seq`, `WCS`, `RAD-Seq`, `CLONE`, `AMPLICON`, `POOLCLONE`, `etc`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | +| `insert_size` | No | Fragment/insert size for paired-end reads (e.g., 500 for 500 bp inserts). Leave empty if not applicable. | +| `library_name` | No | Descriptive library name (optional). | +| `description` | No | Free-text description of the experiment (optional). | ## Submission study From c11bab24fa76bdc982905fc41da18f94fd0ba875 Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Fri, 10 Jul 2026 12:13:14 +0100 Subject: [PATCH 17/47] more fixes --- conf/test_assembly_complete_metadata.config | 2 +- conf/test_mag_complete_metadata.conf | 2 +- docs/usage.md | 58 +++++++++---------- modules/local/count_rna/main.nf | 8 ++- modules/local/count_rna/meta.yml | 4 -- .../create_assembly_metadata_csv/meta.yml | 5 -- .../local/create_genome_metadata_tsv/meta.yml | 5 -- modules/local/ena_webin_cli_wrapper/main.nf | 12 ++++ modules/local/ena_webin_cli_wrapper/meta.yml | 5 -- .../local/generate_assembly_manifest/meta.yml | 4 -- modules/local/genome_upload/meta.yml | 4 -- modules/local/registerstudy/meta.yml | 5 -- .../local/rename_fasta_for_catpack/main.nf | 42 ++++++++------ nextflow_schema.json | 16 ++--- subworkflows/local/genome_evaluation/main.nf | 1 - subworkflows/local/rna_detection/main.nf | 2 - subworkflows/local/rna_detection/meta.yml | 5 -- .../utils_nfcore_seqsubmit_pipeline/main.nf | 5 +- workflows/assemblysubmit.nf | 4 +- workflows/genomesubmit.nf | 19 +++--- 20 files changed, 94 insertions(+), 114 deletions(-) diff --git a/conf/test_assembly_complete_metadata.config b/conf/test_assembly_complete_metadata.config index 16e4ccf..aa8c4d9 100644 --- a/conf/test_assembly_complete_metadata.config +++ b/conf/test_assembly_complete_metadata.config @@ -20,7 +20,7 @@ process { params { config_profile_name = 'Test --mode metagenomic_assemblies complete_metadata profile' - config_profile_description = 'Single-case assembly test with complete metadata values provided' + config_profile_description = 'Single-case assembly test with complete metadata in samplesheet. No statistics calculations will be performed; the pipeline will only upload the data. Study for submission was already registered and provided via --submission_study.' // Input data input = params.pipelines_testdata_base_path + 'seqsubmit/samplesheets/assembly_complete_metadata.csv' diff --git a/conf/test_mag_complete_metadata.conf b/conf/test_mag_complete_metadata.conf index a25e31b..e2d2489 100644 --- a/conf/test_mag_complete_metadata.conf +++ b/conf/test_mag_complete_metadata.conf @@ -20,7 +20,7 @@ process { params { config_profile_name = 'Test --mode mags complete_metadata profile' - config_profile_description = 'Single-case MAG test with complete metadata values provided' + config_profile_description = 'Single-case MAG test with complete metadata in samplesheet. No statistics or taxonomy calculations will be performed; the pipeline will only upload the data. Study for submission was already registered and provided via --submission_study.' // Input data input = params.pipelines_testdata_base_path + 'seqsubmit/samplesheets/mag_complete_metadata.csv' diff --git a/docs/usage.md b/docs/usage.md index 00274a4..756daa1 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -51,27 +51,27 @@ mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default, > [!IMPORTANT] > **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. -| Column | Description | -| --------------------------- | ---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | A unique identifier for this data entry. Must be globally unique within the input dataset. | -| `fasta` | Path to MAG/bin contigs in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. | -| `accession` | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | -| `fastq_1` | Path to the read file in FASTQ format used to generate the source metagenomic assembly. Required if `genome_coverage` is not provided. | -| `fastq_2` | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | -| `assembly_software` | Tool name and version that were used to generate the source metagenomic assembly. | -| `binning_software` | Binning tool, including version, that was used to generate the bins. | -| `binning_parameters` | Arguments that were used during binning. | -| `stats_generation_software` | Tool, including version, that was used to calculate completeness and contamination. | -| `completeness` | Genome completeness value. | -| `contamination` | Genome contamination value. | -| `genome_coverage` | Estimated average sequencing depth across the genome. If the value is missing, it is computed automatically during pipeline execution when reads are provided. | -| `metagenome` | Registered metagenome taxonomic identifier or name that matches an existing ENA taxonomy entry. For more details see https://ena-docs.readthedocs.io/en/latest/faq/taxonomy.html | -| `co-assembly` | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: Yes or No. | -| `broad_environment` | Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428) | -| `local_environment` | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". | +| Column | Description | +| --------------------------- | ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `sample` | A unique identifier for this data entry. Must be globally unique within the input dataset. | +| `fasta` | Path to MAG/bin contigs in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. | +| `accession` | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | +| `fastq_1` | Path to the read file in FASTQ format used to generate the source metagenomic assembly. Required if `genome_coverage` is not provided. | +| `fastq_2` | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | +| `assembly_software` | Tool name and version that were used to generate the source metagenomic assembly. | +| `binning_software` | Binning tool, including version, that was used to generate the bins. | +| `binning_parameters` | Arguments that were used during binning. | +| `stats_generation_software` | Tool, including version, that was used to calculate completeness and contamination. | +| `completeness` | Genome completeness value. | +| `contamination` | Genome contamination value. | +| `genome_coverage` | Estimated average sequencing depth across the genome. If the value is missing, it is computed automatically during pipeline execution when reads are provided. | +| `metagenome` | Registered metagenome taxonomic identifier or name that matches an existing ENA taxonomy entry. For more details see https://ena-docs.readthedocs.io/en/latest/faq/taxonomy.html | +| `co-assembly` | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: Yes or No. | +| `broad_environment` | Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428) | +| `local_environment` | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". | | `environmental_medium` | Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). | -| `RNA_presence` | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: Yes or No. | -| `NCBI_lineage` | NCBI taxonomy lineage of the genome. Can be composted of either numerical IDs or taxon names separated by ";". | +| `RNA_presence` | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: Yes or No. | +| `NCBI_lineage` | NCBI taxonomy lineage of the genome. Can be composted of either numerical IDs or taxon names separated by ";". | > [!NOTE] > More information about envioronment tags can be found at checklists [ERC000050](https://www.ebi.ac.uk/ena/browser/view/ERC000050) for bins and [ERC000047](https://www.ebi.ac.uk/ena/browser/view/ERC000047) for MAGs under the field names "broad-scale environmental context", "local environmental context", and "environmental medium". @@ -93,16 +93,16 @@ assembly_002,data/assembly_002.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 > [!IMPORTANT] > **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. -| Column | Description | -| ------------------- | ---------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | A unique identifier for this data entry. Must be globally unique within the input dataset. | -| `fasta` | Path to assembly contigs in FASTA format compressed with `gzip`. | -| `fastq_1` | Path to the read file in FASTQ format used to generate the metagenomic assembly. Required if `coverage` is not provided. | -| `fastq_2` | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | +| Column | Description | +| ------------------- | ----------------------------------------------------------------------------------------------------------------------------------------------------- | +| `sample` | A unique identifier for this data entry. Must be globally unique within the input dataset. | +| `fasta` | Path to assembly contigs in FASTA format compressed with `gzip`. | +| `fastq_1` | Path to the read file in FASTQ format used to generate the metagenomic assembly. Required if `coverage` is not provided. | +| `fastq_2` | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | | `coverage` | Estimated sequencing depth of the assembly. If this value is missing, it is computed automatically during pipeline execution when reads are provided. | -| `run_accession` | ENA run accession for the reads used to generate the metagenomic assembly. Reads must already be submitted to ENA. | -| `assembler` | Name of the assembler software used to generate the assembly. | -| `assembler_version` | Version of the assembler software used to generate the assembly. | +| `run_accession` | ENA run accession for the reads used to generate the metagenomic assembly. Reads must already be submitted to ENA. | +| `assembler` | Name of the assembler software used to generate the assembly. | +| `assembler_version` | Version of the assembler software used to generate the assembly. | An example file is available at [assets/samplesheet_assembly.csv](../assets/samplesheet_assembly.csv). diff --git a/modules/local/count_rna/main.nf b/modules/local/count_rna/main.nf index b2594b7..2bf5758 100644 --- a/modules/local/count_rna/main.nf +++ b/modules/local/count_rna/main.nf @@ -20,15 +20,19 @@ process COUNT_RNA { tuple val(meta), path("*rna_decision.tsv"), emit: rna_decision tuple val("${task.process}"), val('python'), eval('python --version 2>&1 | sed "s/Python //g"'), topic: versions + when: + task.ext.when == null || task.ext.when + script: + def prefix = task.ext.prefix ?: "${meta.id}" """ count_rna.py \\ --trna ${trnas_stats} \\ --rrna ${rrna_gff} \\ - --name ${meta.id} \\ + --name ${prefix} \\ --trna-limit ${min_trna_count} \\ --rrna-limit ${min_rrna_percentage} \\ - --output ${meta.id}_rna_decision.tsv + --output ${prefix}_rna_decision.tsv """ stub: diff --git a/modules/local/count_rna/meta.yml b/modules/local/count_rna/meta.yml index d7e92f9..4b505cf 100644 --- a/modules/local/count_rna/meta.yml +++ b/modules/local/count_rna/meta.yml @@ -49,10 +49,6 @@ output: Tab-separated file containing genome ID and decision (Yes/No) on whether sufficient RNA genes are present pattern: "*_rna_decision.tsv" - - versions: - type: file - description: File containing software versions - pattern: "versions.yml" authors: - "@KateSakharova" diff --git a/modules/local/create_assembly_metadata_csv/meta.yml b/modules/local/create_assembly_metadata_csv/meta.yml index b1301c7..ffe7be4 100644 --- a/modules/local/create_assembly_metadata_csv/meta.yml +++ b/modules/local/create_assembly_metadata_csv/meta.yml @@ -36,11 +36,6 @@ output: type: file description: CSV file containing assembly metadata to be used as input for the assembly_uploader pattern: "*_assembly_metadata.csv" - - versions: - - versions.yml: - type: file - description: File containing software versions - pattern: "versions.yml" authors: - "@ochkalova" diff --git a/modules/local/create_genome_metadata_tsv/meta.yml b/modules/local/create_genome_metadata_tsv/meta.yml index 74752ad..83f79d3 100644 --- a/modules/local/create_genome_metadata_tsv/meta.yml +++ b/modules/local/create_genome_metadata_tsv/meta.yml @@ -48,11 +48,6 @@ output: type: file description: Tab-separated TSV file with genome metadata pattern: "*.tsv" - - versions: - - versions.yml: - type: file - description: File containing software versions - pattern: "versions.yml" authors: - "@ochkalova" diff --git a/modules/local/ena_webin_cli_wrapper/main.nf b/modules/local/ena_webin_cli_wrapper/main.nf index 6d62fda..7a68961 100644 --- a/modules/local/ena_webin_cli_wrapper/main.nf +++ b/modules/local/ena_webin_cli_wrapper/main.nf @@ -26,6 +26,9 @@ process ENA_WEBIN_CLI_WRAPPER { tuple val("${task.process}"), val('ena-webin-cli'), eval('ena-webin-cli -version'), topic: versions tuple val("${task.process}"), val('mgnify-pipelines-toolkit'), eval('python -c "import importlib.metadata; print(importlib.metadata.version(\'mgnify-pipelines-toolkit\'))"'), topic: versions + when: + task.ext.when == null || task.ext.when + script: def args = task.ext.args ?: "" def prefix = task.ext.prefix ?: "${meta.id}" @@ -40,4 +43,13 @@ process ENA_WEBIN_CLI_WRAPPER { ${test_flag} \\ ${args} """ + + stub: + def prefix = task.ext.prefix ?: "${meta.id}" + if (webincli_mode == "submit") { + """ + touch ${prefix}_accessions.tsv + """ + } + // there is no file for mode=validate } diff --git a/modules/local/ena_webin_cli_wrapper/meta.yml b/modules/local/ena_webin_cli_wrapper/meta.yml index e31a110..62deb91 100644 --- a/modules/local/ena_webin_cli_wrapper/meta.yml +++ b/modules/local/ena_webin_cli_wrapper/meta.yml @@ -47,11 +47,6 @@ output: TSV file containing the accession assigned by ENA for the submitted item. File has two columns: "alias" and "accession". pattern: "*_accessions.tsv" - - versions: - - "versions.yml": - type: file - description: File containing software versions. - pattern: "versions.yml" authors: - "@KateSakharova" diff --git a/modules/local/generate_assembly_manifest/meta.yml b/modules/local/generate_assembly_manifest/meta.yml index 6bd102b..1f9e1de 100644 --- a/modules/local/generate_assembly_manifest/meta.yml +++ b/modules/local/generate_assembly_manifest/meta.yml @@ -50,10 +50,6 @@ output: - manifest: type: file description: Manifest file required for submission with webin-cli. - - versions: - type: file - description: File containing software versions. - pattern: "versions.yml" authors: - "@fxquah" diff --git a/modules/local/genome_upload/meta.yml b/modules/local/genome_upload/meta.yml index 14b5412..4364cf2 100644 --- a/modules/local/genome_upload/meta.yml +++ b/modules/local/genome_upload/meta.yml @@ -61,10 +61,6 @@ output: type: file description: XML file required for ENA submission pattern: "*submission.xml" - - versions: - type: file - description: File containing software versions - pattern: "versions.yml" authors: - "@KateSakharova" diff --git a/modules/local/registerstudy/meta.yml b/modules/local/registerstudy/meta.yml index 803fc62..f4ca931 100644 --- a/modules/local/registerstudy/meta.yml +++ b/modules/local/registerstudy/meta.yml @@ -59,11 +59,6 @@ output: JSON file containing the submission results with keys: submitted (newly created accessions) and failed. pattern: "*_accessions.json" - - versions: - - "versions.yml": - type: file - description: File containing software versions - pattern: "versions.yml" authors: - "@timrozday" diff --git a/modules/local/rename_fasta_for_catpack/main.nf b/modules/local/rename_fasta_for_catpack/main.nf index 38ac856..e70cd15 100644 --- a/modules/local/rename_fasta_for_catpack/main.nf +++ b/modules/local/rename_fasta_for_catpack/main.nf @@ -13,24 +13,32 @@ process RENAME_FASTA_FOR_CATPACK { tuple val(meta), path("output/*.fasta"), emit: renamed_fasta tuple val("${task.process}"), val('bash'), eval('bash --version | head -n1 | sed "s/.*version //; s/ .*//"'), topic: versions + when: + task.ext.when == null || task.ext.when script: - def is_compressed = fasta.name.endsWith('.gz') - extension = '.fasta' - def base = fasta.name - .replaceAll(/\.gz$/, '') - .replaceAll(/\.(fa|fasta|fna)$/, '') - def output_name = base + extension + def is_compressed = fasta.name.endsWith('.gz') + extension = '.fasta' + def base = fasta.name + .replaceAll(/\.gz$/, '') + .replaceAll(/\.(fa|fasta|fna)$/, '') + def output_name = base + extension - if (is_compressed) { - """ - mkdir -p output - gunzip -c ${fasta} > output/${output_name} - """ - } else { - """ - mkdir -p output - ln -s ../${fasta} output/${output_name} - """ - } + if (is_compressed) { + """ + mkdir -p output + gunzip -c ${fasta} > output/${output_name} + """ + } else { + """ + mkdir -p output + ln -s ../${fasta} output/${output_name} + """ + } + + stub: + """ + mkdir -p output + touch output/${meta.id}.fasta + """ } diff --git a/nextflow_schema.json b/nextflow_schema.json index 59c6dc0..1548095 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -57,7 +57,7 @@ }, "trna_limit": { "type": "number", - "description": "Minimum number of tRNA.", + "description": "Minimum number of tRNA required to pass.", "default": 18, "fa_icon": "fas fa-users-cog" } @@ -71,9 +71,9 @@ "help_text": "Provide a path to pre-downloaded DB or DB id for automatic download", "properties": { "cat_db": { - "type": "string", + "type": "file-path", "description": "Path to local CAT_pack DB", - "help": "Database should be pre-downloaded using CATpack commands. The folder with database should contain sub-folders 'db' with files .dmnd, .fastaid2LCAtaxid, .taxids_with_multiple_offspring, etc. And subfolder 'tax' that should contain taxonomy files like .dmp, .taxdump.tar.gz, .prot.accession2taxid.FULL.gz, etc", + "help": "Database should be pre-downloaded using CATpack commands. The folder with database should contain sub-folders 'db' with files .dmnd, .fastaid2LCAtaxid, .taxids_with_multiple_offspring, etc, as well as subfolder 'tax' that should contain taxonomy files like .dmp, .taxdump.tar.gz, .prot.accession2taxid.FULL.gz, etc", "fa_icon": "fas fa-users-cog" }, "cat_db_download_id": { @@ -93,7 +93,7 @@ "help_text": "These options are used to run Checkm2 that will calculate completeness and contamination of genome if those were not provided as input", "properties": { "checkm2_db": { - "type": "string", + "type": "file-path", "description": "Path to pre-downloaded checkm2 database", "fa_icon": "fas fa-users-cog" }, @@ -268,7 +268,7 @@ }, "upload_tpa": { "type": "boolean", - "description": "Flag to control the type of assembly study (third party assembly or not)", + "description": "Flag to control the type of assembly study (third party assembly (TPA) or not). TPA description: https://www.ncbi.nlm.nih.gov/genbank/tpa/.", "default": false, "help": "Use this flag if the study is a third party assembly (TPA). Default: false" }, @@ -289,13 +289,13 @@ "type": "boolean", "description": "Use that flag if you are referring to private data in ENA", "default": false, - "help": "This flag is used during metadata fetch steps. If you are referring to private data (project/reads/assembly) set to True. That flag does NOT regulate privacy of your submission data (check --release_date)" + "help": "This flag is used during metadata fetch steps. Specify if you are referring to private data (project/reads/assembly). That flag does NOT regulate privacy of your submission data (check --release_date)" }, "release_date": { "type": "string", "pattern": "^\\d{4}-\\d{2}-\\d{2}$", "description": "Use that flag if you want to keep your data private after submission", - "help": "This flag regulates privacy of created study for submission. If you want to keep your data private you need to specify a DATE when it can become public (YYYY-MM-DD, max 2 years from now)" + "help": "This flag regulates when the created study for submission will become publically available and indexed on the ENA. If you want to keep your data private until a later date, you need to specify a DATE when it can become public (YYYY-MM-DD, max 2 years from now)" }, "submission_study": { "type": "string", @@ -307,7 +307,7 @@ "format": "file-path", "exists": true, "description": "Path to study metadata file (JSON, CSV, or TSV) for registering a new ENA study. Required when submission_study is not provided.", - "help_text": "File containing study metadata fields (required: study_title and alias, optional: study_abstract, existing_study_type, etc.). Used by REGISTERSTUDY to create a new study in ENA when no existing submission_study accession is given.", + "help_text": "File containing study metadata fields (required: study_title and alias, optional: study_abstract, existing_study_type, etc.). Used by REGISTERSTUDY to create a new study in ENA when no existing submission_study accession is given. ENA documentation: https://ena-docs.readthedocs.io/en/latest/submit/study/programmatic.html", "fa_icon": "fas fa-file-alt" }, "webincli_mode": { diff --git a/subworkflows/local/genome_evaluation/main.nf b/subworkflows/local/genome_evaluation/main.nf index 758f903..4f4921f 100644 --- a/subworkflows/local/genome_evaluation/main.nf +++ b/subworkflows/local/genome_evaluation/main.nf @@ -27,7 +27,6 @@ workflow GENOME_EVALUATION { // only used if ch_checkm2_db is empty main: - ch_versions = channel.empty() // // Database preparation diff --git a/subworkflows/local/rna_detection/main.nf b/subworkflows/local/rna_detection/main.nf index bb0a23f..0cac5bb 100644 --- a/subworkflows/local/rna_detection/main.nf +++ b/subworkflows/local/rna_detection/main.nf @@ -26,7 +26,6 @@ workflow RNA_DETECTION { main: - ch_versions = channel.empty() BARRNAP( fasta.map {id, fasta_file -> [id, fasta_file, "bac"]} ) @@ -43,5 +42,4 @@ workflow RNA_DETECTION { emit: rna_detected = COUNT_RNA.out.rna_decision - versions = ch_versions } diff --git a/subworkflows/local/rna_detection/meta.yml b/subworkflows/local/rna_detection/meta.yml index 4650e92..84902ba 100644 --- a/subworkflows/local/rna_detection/meta.yml +++ b/subworkflows/local/rna_detection/meta.yml @@ -43,11 +43,6 @@ output: pattern: "*.tsv" ontologies: - edam: "http://edamontology.org/format_3475" # TSV - - versions: - type: file - description: | - Channel containing software versions. - Structure: versions authors: - "@KateSakharova" - "@ochkalova" diff --git a/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf b/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf index 53fd0dc..f65014a 100644 --- a/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf @@ -112,10 +112,7 @@ workflow PIPELINE_INITIALISATION { } else if ( mode == "reads" ) { ch_samplesheet = channel .fromList(samplesheetToList(input, "${projectDir}/assets/schema_input_reads.json")) - } else { - error("Unknown mode specified: '${mode}'. Supported modes are 'mags', 'bins', 'metagenomic_assemblies', and 'reads'.") } - emit: samplesheet = ch_samplesheet versions = ch_versions @@ -200,7 +197,7 @@ def toolCitationText() { ].join(' ').trim() def preprocessing_tools = [ - "Input FASTA validation was done by py_fasta_validator (Edwards et al. 2023)." + "Input FASTA validation was done by fa-lint (https://github.com/GallVp/fa-lint)." ].join(' ').trim() def stats_tools = [ diff --git a/workflows/assemblysubmit.nf b/workflows/assemblysubmit.nf index f712b72..d599b59 100644 --- a/workflows/assemblysubmit.nf +++ b/workflows/assemblysubmit.nf @@ -231,8 +231,8 @@ workflow ASSEMBLYSUBMIT { : file("${projectDir}/assets/methods_description_template.yml", checkIfExists: true) def ch_methods_description = channel.value(methodsDescriptionText(ch_multiqc_custom_methods_description)) ch_multiqc_files = ch_multiqc_files.mix(ch_methods_description.collectFile(name: 'methods_description_mqc.yaml', sort: true)) - ch_multiqc_files = ch_multiqc_files.mix(CONCAT_ACCESSIONS.out.file_out.map{meta, file -> file}) - ch_multiqc_files = ch_multiqc_files.mix(CONCAT_METADATA.out.file_out.map{meta, file -> file}) + ch_multiqc_files = ch_multiqc_files.mix(CONCAT_ACCESSIONS.out.file_out.map{_meta, file -> file}) + ch_multiqc_files = ch_multiqc_files.mix(CONCAT_METADATA.out.file_out.map{_meta, file -> file}) MULTIQC( ch_multiqc_files.flatten().collect().map { files -> [ diff --git a/workflows/genomesubmit.nf b/workflows/genomesubmit.nf index 8c1df8e..780a88e 100644 --- a/workflows/genomesubmit.nf +++ b/workflows/genomesubmit.nf @@ -150,7 +150,6 @@ workflow GENOMESUBMIT { trna_limit, rrna_limit ) - ch_versions = ch_versions.mix( RNA_DETECTION.out.versions ) // Update metadata for records missing RNA fasta_updated_with_rna = RNA_DETECTION.out.rna_detected.join(branched_rna_results.rna_prediction_input) @@ -304,13 +303,13 @@ workflow GENOMESUBMIT { } // Combine fasta and manifests ch_combined = fasta_updated_with_taxonomy - .map { meta, fasta -> [meta.id, meta, fasta] } - .join( - manifests_ch.map { meta, manifest -> [meta.id, manifest] } // Has only [id: prefix] - ) - .map { _id, full_meta, fasta, manifest -> - [full_meta, fasta, manifest] - } + .map { meta, fasta -> [meta.id, meta, fasta] } + .join( + manifests_ch.map { meta, manifest -> [meta.id, manifest] } // Has only [id: prefix] + ) + .map { _id, full_meta, fasta, manifest -> + [full_meta, fasta, manifest] + } // --------- Upload data to ENA @@ -369,9 +368,9 @@ workflow GENOMESUBMIT { : file("${projectDir}/assets/methods_description_template.yml", checkIfExists: true) def ch_methods_description = channel.value(methodsDescriptionText(ch_multiqc_custom_methods_description)) ch_multiqc_files = ch_multiqc_files.mix(ch_methods_description.collectFile(name: 'methods_description_mqc.yaml', sort: true)) - ch_multiqc_files = ch_multiqc_files.mix(CONCAT_METADATA.out.file_out.map{meta, file -> file}) + ch_multiqc_files = ch_multiqc_files.mix(CONCAT_METADATA.out.file_out.map{_meta, file -> file}) ch_multiqc_files = ch_multiqc_files.mix(CREATE_MANIFESTS.out.upload_registered_mags) - ch_multiqc_files = ch_multiqc_files.mix(CONCAT_ACCESSIONS.out.file_out.map{meta, file -> file}) + ch_multiqc_files = ch_multiqc_files.mix(CONCAT_ACCESSIONS.out.file_out.map{_meta, file -> file}) MULTIQC( ch_multiqc_files.flatten().collect().map { files -> [ From 6aea0f85bc13d2b711b0e760c6ab220b7e318fd7 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Fri, 10 Jul 2026 13:07:11 +0100 Subject: [PATCH 18/47] slim Samplesheet descriptions in README to minimum --- README.md | 93 +++++---------------------------------------------- docs/usage.md | 6 ++-- 2 files changed, 11 insertions(+), 88 deletions(-) diff --git a/README.md b/README.md index 1a8fd84..f13b535 100644 --- a/README.md +++ b/README.md @@ -65,42 +65,9 @@ Make sure you update commands above with your authorised credentials. ## Input samplesheets -For detailed descriptions of all samplesheet columns, see the [usage documentation](docs/usage.md#samplesheet-input). +### `mags` and `bins` modes -### `mags` and `bins` modes (`GENOMESUBMIT`) - -The input must follow `assets/schema_input_genome.json`. - -Required columns: - -- `sample` -- `fasta` (must end with `.fa.gz`, `.fasta.gz`, or `.fna.gz`) -- `accession` -- `assembly_software` -- `binning_software` -- `binning_parameters` -- `metagenome` -- `environmental_medium` -- `broad_environment` -- `local_environment` -- `co-assembly` - -At least one of the following must be provided per row: - -- reads (`fastq_1`, optional `fastq_2` for paired-end) -- `genome_coverage` - -Additional supported columns: - -- `stats_generation_software` -- `completeness` -- `contamination` -- `RNA_presence` -- `NCBI_lineage` - -If `genome_coverage`, `stats_generation_software`, `completeness`, `contamination`, `RNA_presence`, or `NCBI_lineage` are missing, the workflow can calculate or infer them when the required inputs are available. See the [Methods documentation](docs/methods.md) for more information on how metadata statistics are obtained. - -Those fields are metadata required for the [genome_uploader](https://github.com/EBI-Metagenomics/genome_uploader) package. +The input must follow [assets/schema_input_genome.json](assets/schema_input_genome.json). See the [corresponding samplesheet section](docs/usage.md#mags-and-bins-modes-genomesubmit) of the usage documentation for the full list of required and optional columns. Example `samplesheet_genomes.csv`: @@ -109,27 +76,9 @@ sample,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binnin lachnospira_eligens,data/bin_lachnospira_eligens.fa.gz,SRR24458089,,,spades_v3.15.5,metabat2_v2.6,default,CheckM2_v1.0.1,61.0,0.21,32.07,sediment metagenome,No,marine,cable_bacteria,marine_sediment,No,d__Bacteria;p__Proteobacteria;s__unclassified_Proteobacteria ``` -> [!IMPORTANT] -> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. - -### `metagenomic_assemblies` mode (`ASSEMBLYSUBMIT`) - -The input must follow `assets/schema_input_assembly.json`. +### `metagenomic_assemblies` mode -Required columns: - -- `sample` -- `fasta` (must end with `.fa.gz`, `.fasta.gz`, or `.fna.gz`) -- `run_accession` -- `assembler` -- `assembler_version` - -At least one of the following must be provided per row: - -- reads (`fastq_1`, optional `fastq_2` for paired-end) -- `coverage` - -If `coverage` is missing and reads are provided, the workflow calculates average coverage with `coverm`. See the [Methods documentation](docs/methods.md) for more information on how coverage is calculated. +The input must follow [assets/schema_input_assembly.json](assets/schema_input_assembly.json). See the [corresponding samplesheet section](docs/usage.md#metagenomic_assemblies-mode-assemblysubmit) of the usage documentation for more details. Example `samplesheet_assembly.csv`: @@ -139,30 +88,9 @@ assembly_1,data/contigs_1.fasta.gz,data/reads_1.fastq.gz,data/reads_2.fastq.gz,, assembly_2,data/contigs_2.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 ``` -> [!IMPORTANT] -> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. - -### `reads` mode (`READSUBMIT`) - -The input must follow `assets/schema_input_reads.json`. +### `reads` mode -Required columns: - -- `sample` -- `sample_accession` -- `fastq_1` -- `fastq_2` -- `platform` -- `instrument` -- `library_source` -- `library_selection` -- `library_strategy` - -Optional columns: - -- `insert_size` -- `library_name` -- `description` +The input must follow [assets/schema_input_reads.json](assets/schema_input_reads.json). See the [corresponding samplesheet section](docs/usage.md#reads-mode-readsubmit) of the usage documentation for more details. Example `samplesheet_reads.csv`: @@ -171,9 +99,6 @@ sample,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,libra illumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLUMINA,Illumina HiSeq 2000,GENOMIC,RANDOM,WGS,500,HiSeq_library_001,Illumina sequencing of sample XYZ ``` -> [!IMPORTANT] -> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. - ## Usage > [!NOTE] @@ -181,9 +106,7 @@ illumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLU ### Submission study -All data submitted through this pipeline must be associated with an ENA study (project). You can either pass an accession of your existing study via `--submission_study`or provide a metadata file via `--study_metadata` and the pipeline will register the study with ENA before submitting your data. - -See the [usage documentation](docs/usage.md#submission-study) for more details. +All data submitted through this pipeline must be associated with an ENA study (project). You can either pass an accession of your existing study via `--submission_study`or provide a metadata file via `--study_metadata` and the pipeline will register the study with ENA before submitting your data. See the [usage documentation](docs/usage.md#submission-study) for more details. ### Data privacy @@ -191,7 +114,7 @@ The pipeline supports private submissions. See the [usage documentation](docs/us ### Database setup (`CheckM2` and `CAT_pack`) -The `mags`/`bins` workflow requires databases for completeness/contamination estimation and taxonomy assignment. See [Usage documentation](usage.md) for details. +The `mags`/`bins` workflow requires databases for completeness/contamination estimation and taxonomy assignment. See [Usage documentation](usage.md) for more details. ### Required parameters: diff --git a/docs/usage.md b/docs/usage.md index bc30044..d666aad 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -76,12 +76,12 @@ mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default, > [!NOTE] > More information about envioronment tags can be found at checklists [ERC000050](https://www.ebi.ac.uk/ena/browser/view/ERC000050) for bins and [ERC000047](https://www.ebi.ac.uk/ena/browser/view/ERC000047) for MAGs under the field names "broad-scale environmental context", "local environmental context", and "environmental medium". +If `genome_coverage`, `stats_generation_software`, `completeness`, `contamination`, `RNA_presence`, or `NCBI_lineage` are missing, the workflow can calculate or infer them when the required inputs are available. See the [Methods documentation](docs/methods.md) for more information on how metadata statistics are obtained. + ### `metagenomic_assemblies` mode (`ASSEMBLYSUBMIT`) Use this samplesheet structure for metagenomic assembly submission. The input format follows [assets/schema_input_assembly.json](../assets/schema_input_assembly.json). -Provide either read files (`fastq_1`, optionally `fastq_2`) or a `coverage` value for each row. If `coverage` is missing and reads are provided, the workflow calculates average coverage automatically. - Example: ```csv title="samplesheet_assembly.csv" @@ -104,7 +104,7 @@ assembly_002,data/assembly_002.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 | `assembler` | Yes | Name of the assembler software used to generate the assembly. | | `assembler_version` | Yes | Version of the assembler software used to generate the assembly. | -An example file is available at [assets/samplesheet_assembly.csv](../assets/samplesheet_assembly.csv). +Provide either read files (`fastq_1`, optionally `fastq_2`) or a `coverage` value for each row. If `coverage` is missing and reads are provided, the workflow calculates average coverage with `coverm`. See the [Methods documentation](docs/methods.md) for more information on how coverage is calculated. ### `reads` mode (`READSUBMIT`) From 93940733a3fea8ff3757c12158e985bfc0ec9af5 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Fri, 10 Jul 2026 13:49:34 +0100 Subject: [PATCH 19/47] Add Table of Contents to usage.md --- docs/usage.md | 17 +++++++++++++++++ 1 file changed, 17 insertions(+) diff --git a/docs/usage.md b/docs/usage.md index d666aad..995ec2a 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -4,6 +4,23 @@ > _Documentation of pipeline parameters is generated automatically from the pipeline schema and can no longer be found in markdown files._ +## Table of Contents + +- [Introduction](#introduction) +- [Before you start](#before-you-start) +- [Samplesheet input](#samplesheet-input) + - [`mags` and `bins` modes](#mags-and-bins-modes-genomesubmit) + - [`metagenomic_assemblies` mode](#metagenomic_assemblies-mode-assemblysubmit) + - [`reads` mode](#reads-mode-readsubmit) +- [Submission study](#submission-study) +- [Data privacy](#data-privacy) +- [Database preparation (`mags` / `bins` modes)](#database-preparation-mags--bins) +- [Running the pipeline](#running-the-pipeline) +- [Core Nextflow arguments](#core-nextflow-arguments) +- [Custom configuration](#custom-configuration) +- [Running in the background](#running-in-the-background) +- [Nextflow memory requirements](#nextflow-memory-requirements) + ## Introduction `nf-core/seqsubmit` is a Nextflow pipeline for submitting metagenomic assemblies, MAGs, bins, and raw reads to ENA. From cd8d99a23b7b76ce70fde5c03533de786d3ad7e0 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Fri, 10 Jul 2026 13:54:33 +0100 Subject: [PATCH 20/47] move Submission Study section up in usage.md --- docs/usage.md | 126 +++++++++++++++++++++++++------------------------- 1 file changed, 63 insertions(+), 63 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index 995ec2a..eaef351 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -8,11 +8,11 @@ - [Introduction](#introduction) - [Before you start](#before-you-start) +- [Submission study](#submission-study) - [Samplesheet input](#samplesheet-input) - [`mags` and `bins` modes](#mags-and-bins-modes-genomesubmit) - [`metagenomic_assemblies` mode](#metagenomic_assemblies-mode-assemblysubmit) - [`reads` mode](#reads-mode-readsubmit) -- [Submission study](#submission-study) - [Data privacy](#data-privacy) - [Database preparation (`mags` / `bins` modes)](#database-preparation-mags--bins) - [Running the pipeline](#running-the-pipeline) @@ -46,6 +46,68 @@ nextflow secrets set ENA_WEBIN "Webin-XXX" nextflow secrets set ENA_WEBIN_PASSWORD "XXX" ``` +## Submission study + +All data submitted through this pipeline must be associated with an ENA study (project). You have two options: + +### Option 1 — Use an existing study + +If you already have an ENA study, pass its accession (starting with `PRJ` or `ERP`) via `--submission_study`: + +```bash +--submission_study PRJEB12345 +``` + +You can create a study manually via the [Webin Portal](https://www.ebi.ac.uk/ena/submit/webin/login) and then use the assigned accession here. + +### Option 2 — Register a new study automatically + +Provide a study metadata file via `--study_metadata` and the pipeline will register the study with ENA before submitting your data: + +```bash +--study_metadata study_metadata.json +``` + +The pipeline accepts JSON, CSV, and TSV formats. + +#### JSON formats + +Single study as a flat object: + +```json +{ + "alias": "study-gut-2026", + "study_title": "Gut microbiome study", + "study_abstract": "Characterisation of gut microbial communities" +} +``` + +#### CSV format + +```csv +alias,study_title,study_abstract +study-gut-2026,Gut microbiome study,Characterisation of gut microbial communities +``` + +#### TSV format + +```tsv +alias study_title study_abstract +study-soil-2026 Soil microbiome study Survey of soil microbiota +``` + +#### Study metadata fields + +| Field | Required | Description | +| --------------------- | -------- | ---------------------------------------------------------------------------- | +| `study_title` | Yes | Descriptive title of the study. | +| `alias` | Yes | Unique project alias within your Webin account. Max length is 50 characters. | +| `study_abstract` | No | Free-text abstract describing the study. | +| `study_description` | No | Alternative to `study_abstract`. | +| `project_name` | No | Project name. Defaults to `study_title`. | +| `existing_study_type` | No | ENA study type (e.g. `Metagenomics`, `Other`). | +| `new_study_type` | No | Custom study type. Only used when `existing_study_type` is set to `Other`. | + ## Samplesheet input You will need to create a samplesheet with information about the data entries you would like to process before running the pipeline. Use `--input` parameter to specify its location. It has to be a comma-separated file with the structure defined by the execution `--mode`. @@ -153,68 +215,6 @@ pacbio_run_001,SAMEA7654321,data/pacbio_reads.fastq.gz,,PACBIO_SMRT,PacBio Seque | `library_name` | No | Descriptive library name (optional). | | `description` | No | Free-text description of the experiment (optional). | -## Submission study - -All data submitted through this pipeline must be associated with an ENA study (project). You have two options: - -### Option 1 — Use an existing study - -If you already have an ENA study, pass its accession (starting with `PRJ` or `ERP`) via `--submission_study`: - -```bash ---submission_study PRJEB12345 -``` - -You can create a study manually via the [Webin Portal](https://www.ebi.ac.uk/ena/submit/webin/login) and then use the assigned accession here. - -### Option 2 — Register a new study automatically - -Provide a study metadata file via `--study_metadata` and the pipeline will register the study with ENA before submitting your data: - -```bash ---study_metadata study_metadata.json -``` - -The pipeline accepts JSON, CSV, and TSV formats. - -#### JSON formats - -Single study as a flat object: - -```json -{ - "alias": "study-gut-2026", - "study_title": "Gut microbiome study", - "study_abstract": "Characterisation of gut microbial communities" -} -``` - -#### CSV format - -```csv -alias,study_title,study_abstract -study-gut-2026,Gut microbiome study,Characterisation of gut microbial communities -``` - -#### TSV format - -```tsv -alias study_title study_abstract -study-soil-2026 Soil microbiome study Survey of soil microbiota -``` - -#### Study metadata fields - -| Field | Required | Description | -| --------------------- | -------- | ---------------------------------------------------------------------------- | -| `study_title` | Yes | Descriptive title of the study. | -| `alias` | Yes | Unique project alias within your Webin account. Max length is 50 characters. | -| `study_abstract` | No | Free-text abstract describing the study. | -| `study_description` | No | Alternative to `study_abstract`. | -| `project_name` | No | Project name. Defaults to `study_title`. | -| `existing_study_type` | No | ENA study type (e.g. `Metagenomics`, `Other`). | -| `new_study_type` | No | Custom study type. Only used when `existing_study_type` is set to `Other`. | - ## Data privacy You can reference private ENA data if you have access to it via your Webin account (it was submitted previously under your credentials). To use private data accessions in your submission metadata, specify the `--is_private` flag. The pipeline will then use your provided Webin credentials to fetch the required metadata. From ae6b4627c82977799cfe88f844bfd5a251a02ecc Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Fri, 10 Jul 2026 14:14:50 +0100 Subject: [PATCH 21/47] Expand Requirements to include required pre-submissions --- README.md | 6 ++++-- docs/usage.md | 15 +++++++++------ 2 files changed, 13 insertions(+), 8 deletions(-) diff --git a/README.md b/README.md index f13b535..e66d1de 100644 --- a/README.md +++ b/README.md @@ -53,9 +53,11 @@ Currently, the pipeline supports four submission modes, each routed to a dedicat - [Nextflow](https://www.nextflow.io/) `>=25.04.0` - Webin account registered at https://www.ebi.ac.uk/ena/submit/webin/login -- Raw reads used to assemble contigs submitted to [INSDC](https://www.insdc.org/) and associated accessions available +- For metagenomic assemblies submission the raw reads used to generate the assemblies have to be submitted to INSDC/ENA to obtain the corresponding run accessions. You can use the pipeline in mode `reads` to perform this. +- For MAGs or bins submission the raw reads only or raw reads and metagenomic assemblies used to generate the MAGs/bins have to be submitted to INSDC/ENA to obtain the corresponding run or assembly accessions. You can use the pipeline in mode `reads` or `metagenomic_assemblies` to perform this. +- For raw reads submission it is required to register source BioSamples and obtain the sample accessions. -Setup your environment secrets before running the pipeline: +Setup Webin credentials as Nextflow secrets before running the pipeline: `nextflow secrets set ENA_WEBIN "Webin-XXX"` diff --git a/docs/usage.md b/docs/usage.md index eaef351..949e1b4 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -37,14 +37,17 @@ Before running the pipeline, make sure that: - Nextflow `>=25.04.0` is available. - You have a Webin account registered at . -- The raw reads used to generate the submitted assemblies have already been submitted to INSDC/ENA and the relevant accessions are available. +- For metagenomic assemblies submission the raw reads used to generate the assemblies have to be submitted to INSDC/ENA to obtain the corresponding run accessions. You can use the pipeline in mode `reads` to perform this. +- For MAGs or bins submission the raw reads only or raw reads and metagenomic assemblies used to generate the MAGs/bins have to be submitted to INSDC/ENA to obtain the corresponding run or assembly accessions. You can use the pipeline in mode `reads` or `metagenomic_assemblies` to perform this. +- For raw reads submission it is required to register source BioSamples and obtain the sample accessions. -Set your Webin credentials as Nextflow secrets: +Setup Webin credentials as Nextflow secrets before running the pipeline: -```bash -nextflow secrets set ENA_WEBIN "Webin-XXX" -nextflow secrets set ENA_WEBIN_PASSWORD "XXX" -``` +`nextflow secrets set ENA_WEBIN "Webin-XXX"` + +`nextflow secrets set ENA_WEBIN_PASSWORD "XXX"` + +Make sure you update commands above with your authorised credentials. ## Submission study From bea141a3c6b922941b68e823ff2ac93af6d5514e Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Fri, 10 Jul 2026 16:26:42 +0100 Subject: [PATCH 22/47] Apply James' suggesstions in usage.md --- docs/usage.md | 20 +++++++++----------- 1 file changed, 9 insertions(+), 11 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index 949e1b4..ad69cdb 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -153,10 +153,10 @@ mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default, | `local_environment` | Yes | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". | | `environmental_medium` | Yes | Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). | | `RNA_presence` | No | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: Yes or No. | -| `NCBI_lineage` | No | NCBI taxonomy lineage of the genome. Can be composted of either numerical IDs or taxon names separated by ";". | +| `NCBI_lineage` | No | NCBI taxonomy lineage of the genome. Can be composted of either numerical IDs or official NCBI taxon names separated by ";". | > [!NOTE] -> More information about envioronment tags can be found at checklists [ERC000050](https://www.ebi.ac.uk/ena/browser/view/ERC000050) for bins and [ERC000047](https://www.ebi.ac.uk/ena/browser/view/ERC000047) for MAGs under the field names "broad-scale environmental context", "local environmental context", and "environmental medium". +> More information about environment tags can be found at checklists [ERC000050](https://www.ebi.ac.uk/ena/browser/view/ERC000050) for bins and [ERC000047](https://www.ebi.ac.uk/ena/browser/view/ERC000047) for MAGs under the field names "broad-scale environmental context", "local environmental context", and "environmental medium". If `genome_coverage`, `stats_generation_software`, `completeness`, `contamination`, `RNA_presence`, or `NCBI_lineage` are missing, the workflow can calculate or infer them when the required inputs are available. See the [Methods documentation](docs/methods.md) for more information on how metadata statistics are obtained. @@ -210,7 +210,7 @@ pacbio_run_001,SAMEA7654321,data/pacbio_reads.fastq.gz,,PACBIO_SMRT,PacBio Seque | `fastq_1` | Yes | Path to forward reads in FASTQ format (optionally gzipped). | | `fastq_2` | No | Path to reverse reads for paired-end data. Leave empty for single-end reads. | | `platform` | Yes | Sequencing platform. Supported values: `ILLUMINA`, `PACBIO_SMRT`, `OXFORD_NANOPORE`, `ION_TORRENT`, `CAPILLARY`, `DNBSEQ`, `ELEMENT`, `GENAPSYS`, `GENEMIND`, `HELICOS`, `LS454`, `BGISEQ`, `ULTIMA`, `VELA_DIAGNOSTICS`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | -| `instrument` | Yes | Sequencer model, e.g. "Illumina HiSeq 2000", "PacBio Sequel", "MinION". | +| `instrument` | Yes | Sequencer model, e.g. "Illumina HiSeq 2000", "PacBio Sequel", "MinION". See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#permitted-values-for-instrument) | | `library_source` | Yes | Library source type. Options: `GENOMIC`, `METAGENOMIC`, `TRANSCRIPTOMIC`, `METAGENOMIC SINGLE CELL`, `TRANSCRIPTOMIC SINGLE CELL`, `SYNTHETIC`, `VIRAL RNA`, `OTHER`. | | `library_selection` | Yes | Library selection method. Options: `RANDOM`, `PCR`, `RANDOM PCR`, `RT-PCR`, `MF`, `cDNA`, `cDNA_randomPriming`, `cDNA_oligo_dT`, `PolyA`, `Inverse rRNA`, `ChIP`, `MNase`, `DNase`, `Hybrid Selection`, etc. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | | `library_strategy` | Yes | Library strategy. Options: `WGS`, `WGA`, `WXS`, `RNA-Seq`, `miRNA-Seq`, `ncRNA-Seq`, `EST`, `Hi-C`, `ATAC-seq`, `WCS`, `RAD-Seq`, `CLONE`, `AMPLICON`, `POOLCLONE`, `etc`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | @@ -241,21 +241,19 @@ The `GENOMESUBMIT` workflow uses `CheckM2` and `CAT_pack` that require specializ You can either provide pre-existing databases or let the pipeline prepare them during execution. +As databases preparation can take significant time, we strongly recommend downloading them locally and storing them in a local cache folder for reuse across runs. +In particular the `CAT_pack` database is extremely large and may take a long time to download and compute. + - `CheckM2`: - - provide the path to local database with `--checkm2_db`, otherwise the pipeline downloads version specified with `--checkm2_db_download_id` (by default `14897628`). + - provide the path to local database with `--checkm2_db`, otherwise the pipeline downloads version specified with `--checkm2_db_download_id` (by default Zenodo accession `14897628`). - `CAT_pack`: - provide the path to local database (containing `tax/` and `db/` folders or `tar.gz` archive) with `--cat_db`, otherwise the pipeline constructs version specified with `--cat_db_download_id` (by default `nr`). See [CAT_pack documentation](https://github.com/MGXlab/CAT_pack) and [CheckM2 documentation](https://github.com/chklovski/CheckM2) for more details on usage and creation of databases. -> [!IMPORTANT] -> `CAT_pack` database creation can take significant time. -> -> Reusing an existing database is strongly recommended for repeated runs. -> -> Databases created/downloaded by the pipeline are published under: -> `${params.outdir}/databases/` +Databases created/downloaded by the pipeline are published under: +`/databases/`. In the subsequent pipeline runs it is strongly recommended to reuse them using `--cat_db /databases/cat_pack/` and `--cat_db /databases/checkm2/`. ## Running the pipeline From 59c774654cef2c04b30ea4401dd2a654d4a070e9 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Wed, 15 Jul 2026 16:29:22 +0100 Subject: [PATCH 23/47] restructure usage.md --- docs/usage.md | 381 +++++++++++++++++++++++++++++++------------------- 1 file changed, 241 insertions(+), 140 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index ad69cdb..aa43cc0 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -9,13 +9,26 @@ - [Introduction](#introduction) - [Before you start](#before-you-start) - [Submission study](#submission-study) -- [Samplesheet input](#samplesheet-input) - - [`mags` and `bins` modes](#mags-and-bins-modes-genomesubmit) - - [`metagenomic_assemblies` mode](#metagenomic_assemblies-mode-assemblysubmit) - - [`reads` mode](#reads-mode-readsubmit) +- [`reads` mode](#reads-mode) + - [Prerequisites](#prerequisites) + - [Samplesheet input](#samplesheet-input) + - [Running the pipeline in `reads` mode](#running-the-pipeline-in-reads-mode) + - [Limitations](#limitations) +- [`metagenomic_assemblies` mode](#metagenomic_assemblies-mode) + - [Prerequisites](#prerequisites-1) + - [Samplesheet input](#samplesheet-input-1) + - [Running the pipeline in `metagenomic_assemblies` mode](#running-the-pipeline-in-metagenomic_assemblies-mode) + - [Limitations](#limitations-1) +- [`mags` and `bins` modes](#mags-and-bins-modes) + - [Prerequisites](#prerequisites-2) + - [Samplesheet input](#samplesheet-input-2) + - [Database preparation (`CheckM2` and `CAT_pack`)](#database-preparation-checkm2-and-cat_pack) + - [Running the pipeline in `mags` and `bins` modes](#running-the-pipeline-in-mags-and-bins-modes) + - [Limitations](#limitations-2) - [Data privacy](#data-privacy) -- [Database preparation (`mags` / `bins` modes)](#database-preparation-mags--bins) - [Running the pipeline](#running-the-pipeline) + - [Updating the pipeline](#updating-the-pipeline) + - [Reproducibility](#reproducibility) - [Core Nextflow arguments](#core-nextflow-arguments) - [Custom configuration](#custom-configuration) - [Running in the background](#running-in-the-background) @@ -23,31 +36,33 @@ ## Introduction -`nf-core/seqsubmit` is a Nextflow pipeline for submitting metagenomic assemblies, MAGs, bins, and raw reads to ENA. +`nf-core/seqsubmit` is a Nextflow pipeline for submitting sequence data to ENA. -The pipeline supports three workflow paths: +The pipeline supports of the following data types, selected via `--mode` parameter: -- `GENOMESUBMIT` for `--mode mags` and `--mode bins` -- `ASSEMBLYSUBMIT` for `--mode metagenomic_assemblies` -- `READSUBMIT` for `--mode reads` +- `reads` — raw sequencing reads +- `metagenomic_assemblies` — metagenomic assemblies +- `mags` / `bins` — metagenome-assembled genomes (MAGs) or metagenomic bins + +Each mode has its own samplesheet structure, prerequisites, and limitations — see the dedicated section for the mode you want to use below. The [Before you start](#before-you-start), [Submission study](#submission-study), [Running the pipeline](#running-the-pipeline), and [Data privacy](#data-privacy) sections apply to all modes. ## Before you start -Before running the pipeline, make sure that: +Before running the pipeline in any mode, make sure that: - Nextflow `>=25.04.0` is available. - You have a Webin account registered at . -- For metagenomic assemblies submission the raw reads used to generate the assemblies have to be submitted to INSDC/ENA to obtain the corresponding run accessions. You can use the pipeline in mode `reads` to perform this. -- For MAGs or bins submission the raw reads only or raw reads and metagenomic assemblies used to generate the MAGs/bins have to be submitted to INSDC/ENA to obtain the corresponding run or assembly accessions. You can use the pipeline in mode `reads` or `metagenomic_assemblies` to perform this. -- For raw reads submission it is required to register source BioSamples and obtain the sample accessions. -Setup Webin credentials as Nextflow secrets before running the pipeline: + Set your Webin credentials as Nextflow secrets: -`nextflow secrets set ENA_WEBIN "Webin-XXX"` + ```bash + nextflow secrets set ENA_WEBIN "Webin-XXX" + nextflow secrets set ENA_WEBIN_PASSWORD "XXX" + ``` -`nextflow secrets set ENA_WEBIN_PASSWORD "XXX"` +- You prepared either study accession or study metadata file to set a submission study to assosiate the submission with. See [Submission study](#submission-study) section below for details. -Make sure you update commands above with your authorised credentials. +Each mode also has additional prerequisites — see its own "Prerequisites" subsection below. ## Submission study @@ -61,7 +76,7 @@ If you already have an ENA study, pass its accession (starting with `PRJ` or `ER --submission_study PRJEB12345 ``` -You can create a study manually via the [Webin Portal](https://www.ebi.ac.uk/ena/submit/webin/login) and then use the assigned accession here. +You can create a study manually via the [Webin Portal](https://www.ebi.ac.uk/ena/submit/webin/login) following [these instructions](https://ena-docs.readthedocs.io/en/latest/submit/study/interactive.html) and then use the assigned study accession. ### Option 2 — Register a new study automatically @@ -111,58 +126,78 @@ study-soil-2026 Soil microbiome study Survey of soil microbiota | `existing_study_type` | No | ENA study type (e.g. `Metagenomics`, `Other`). | | `new_study_type` | No | Custom study type. Only used when `existing_study_type` is set to `Other`. | -## Samplesheet input +## `reads` mode -You will need to create a samplesheet with information about the data entries you would like to process before running the pipeline. Use `--input` parameter to specify its location. It has to be a comma-separated file with the structure defined by the execution `--mode`. +### Prerequisites -```bash ---input '[path to samplesheet.csv]' -``` +In addition to the general [prerequisites](#before-you-start), for `reads` mode: + +- You need to register the source BioSamples and obtain the sample accessions as described in [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/samples.html). Those have to be provided in `sample_accession` column of the samplesheet. -### `mags` and `bins` modes (`GENOMESUBMIT`) +### Samplesheet input -Use this samplesheet structure for MAG and bin submission. The input format follows [assets/schema_input_genome.json](../assets/schema_input_genome.json). +You will need to create a samplesheet with information about the raw reads you would like to submit before running the pipeline. Use `--input` to specify its location. It must be a comma-separated file with the structure that follows [assets/schema_input_reads.json](../assets/schema_input_reads.json). Example: -```csv title="samplesheet_genomes.csv" -sample,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage -mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default,CheckM2 1.0.1,92.81,1.09,66.04,sediment metagenome,No,marine,cable bacteria,marine sediment,No,d__Bacteria;p__Proteobacteria;s__ +```csv title="samplesheet_reads.csv" +sample,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,library_selection,library_strategy,insert_size,library_name,description +illumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLUMINA,Illumina HiSeq 2000,GENOMIC,RANDOM,WGS,500,HiSeq_library_001,Illumina sequencing of sample XYZ +pacbio_run_001,SAMEA7654321,data/pacbio_reads.fastq.gz,,PACBIO_SMRT,PacBio Sequel,GENOMIC,RANDOM,WGS,,PacBio_library_002,Long-read sequencing ``` > [!IMPORTANT] > **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. -| Column | Required | Description | -| --------------------------- | ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset (used as `meta.id` throughout the pipeline). | -| `fasta` | Yes | Path to MAG/bin contigs in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. | -| `accession` | Yes | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | -| `fastq_1` | Conditional | Path to the read file in FASTQ format used to generate the source metagenomic assembly. Required if `genome_coverage` is not provided. | -| `fastq_2` | No | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | -| `assembly_software` | Yes | Tool name and version that were used to generate the source metagenomic assembly. | -| `binning_software` | Yes | Binning tool, including version, that was used to generate the bins. | -| `binning_parameters` | Yes | Arguments that were used during binning. | -| `stats_generation_software` | No | Tool, including version, that was used to calculate completeness and contamination. | -| `completeness` | No | Genome completeness value. | -| `contamination` | No | Genome contamination value. | -| `genome_coverage` | Conditional | Estimated average sequencing depth across the genome. If the value is missing, it is computed automatically during pipeline execution when reads are provided. | -| `metagenome` | Yes | Registered metagenome taxonomic identifier or name that matches an existing ENA taxonomy entry. For more details see https://ena-docs.readthedocs.io/en/latest/faq/taxonomy.html | -| `co-assembly` | Yes | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: Yes or No. | -| `broad_environment` | Yes | Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428) | -| `local_environment` | Yes | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". | -| `environmental_medium` | Yes | Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). | -| `RNA_presence` | No | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: Yes or No. | -| `NCBI_lineage` | No | NCBI taxonomy lineage of the genome. Can be composted of either numerical IDs or official NCBI taxon names separated by ";". | +| Column | Required | Description | +| ------------------- | -------- | --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `sample` | Yes | Unique identifier of this particular data entry. Used as an experiment name. | +| `sample_accession` | Yes | ENA sample accession (starting with SAMEA) of the sample used to generate raw reads. | +| `fastq_1` | Yes | Path to forward reads in FASTQ format (optionally gzipped). | +| `fastq_2` | No | Path to reverse reads for paired-end data. Leave empty for single-end reads. | +| `platform` | Yes | Sequencing platform. Supported values: `ILLUMINA`, `PACBIO_SMRT`, `OXFORD_NANOPORE`, `ION_TORRENT`, `CAPILLARY`, `DNBSEQ`, `ELEMENT`, `GENAPSYS`, `GENEMIND`, `HELICOS`, `LS454`, `BGISEQ`, `ULTIMA`, `VELA_DIAGNOSTICS`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | +| `instrument` | Yes | Sequencer model, e.g. "Illumina HiSeq 2000", "PacBio Sequel", "MinION". See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#permitted-values-for-instrument). | +| `library_source` | Yes | Library source type. Options: `GENOMIC`, `METAGENOMIC`, `TRANSCRIPTOMIC`, `METAGENOMIC SINGLE CELL`, `TRANSCRIPTOMIC SINGLE CELL`, `SYNTHETIC`, `VIRAL RNA`, `OTHER`. | +| `library_selection` | Yes | Library selection method. Options: `RANDOM`, `PCR`, `RANDOM PCR`, `RT-PCR`, `MF`, `cDNA`, `cDNA_randomPriming`, `cDNA_oligo_dT`, `PolyA`, `Inverse rRNA`, `ChIP`, `MNase`, `DNase`, `Hybrid Selection`, etc. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | +| `library_strategy` | Yes | Library strategy. Options: `WGS`, `WGA`, `WXS`, `RNA-Seq`, `miRNA-Seq`, `ncRNA-Seq`, `EST`, `Hi-C`, `ATAC-seq`, `WCS`, `RAD-Seq`, `CLONE`, `AMPLICON`, `POOLCLONE`, `etc`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | +| `insert_size` | No | Fragment/insert size for paired-end reads (e.g., 500 for 500 bp inserts). Leave empty if not applicable. | +| `library_name` | No | Descriptive library name (optional). | +| `description` | No | Free-text description of the experiment (optional). | -> [!NOTE] -> More information about environment tags can be found at checklists [ERC000050](https://www.ebi.ac.uk/ena/browser/view/ERC000050) for bins and [ERC000047](https://www.ebi.ac.uk/ena/browser/view/ERC000047) for MAGs under the field names "broad-scale environmental context", "local environmental context", and "environmental medium". +### Running the pipeline in `reads` mode -If `genome_coverage`, `stats_generation_software`, `completeness`, `contamination`, `RNA_presence`, or `NCBI_lineage` are missing, the workflow can calculate or infer them when the required inputs are available. See the [Methods documentation](docs/methods.md) for more information on how metadata statistics are obtained. +`reads` mode does not add any parameters beyond the [common parameters](#running-the-pipeline). + +Example `reads` mode upload to TEST server, run command with docker: + +```bash +nextflow run nf-core/seqsubmit \ + -profile docker \ + --mode reads \ + --input samplesheet_reads.csv \ + --submission_study \ + --webincli_mode submit \ + --test_upload \ + --outdir reads_submission/ +``` + +### Limitations + +`reads` mode does not support the following: + +- **Reads in non-FASTQ formats**: Only reads provided as FASTQ (`.fastq.gz`) can be submitted. Other formats (e.g. BAM) are not supported as input. + +## `metagenomic_assemblies` mode + +### Prerequisites -### `metagenomic_assemblies` mode (`ASSEMBLYSUBMIT`) +In addition to the general [prerequisites](#before-you-start), for `metagenomic_assemblies` mode: -Use this samplesheet structure for metagenomic assembly submission. The input format follows [assets/schema_input_assembly.json](../assets/schema_input_assembly.json). +- The raw reads used to generate the assemblies have have to be submitted to INSDC/ENA to obtain the corresponding run accessions. Refer to the [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads.html) for details. You can use `seqsubmit` in `reads` mode to do this. Assigned run accessions have to be provided in `run_accession` column of the samplesheet. + +### Samplesheet input + +You will need to create a samplesheet with information about the metagenomic assemblies you would like to submit before running the pipeline. Use `--input` to specify its location. It must be a comma-separated file with the structure that follows [assets/schema_input_assembly.json](../assets/schema_input_assembly.json). Example: @@ -177,7 +212,7 @@ assembly_002,data/assembly_002.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 | Column | Required | Description | | ------------------- | ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset (used as `meta.id` throughout the pipeline). | +| `sample` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset. | | `fasta` | Yes | Path to assembly contigs in FASTA format compressed with `gzip`. | | `fastq_1` | Conditional | Path to the read file in FASTQ format used to generate the metagenomic assembly. Required if `coverage` is not provided. | | `fastq_2` | No | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | @@ -188,56 +223,104 @@ assembly_002,data/assembly_002.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 Provide either read files (`fastq_1`, optionally `fastq_2`) or a `coverage` value for each row. If `coverage` is missing and reads are provided, the workflow calculates average coverage with `coverm`. See the [Methods documentation](docs/methods.md) for more information on how coverage is calculated. -### `reads` mode (`READSUBMIT`) +### Running the pipeline in `metagenomic_assemblies` mode -Use this samplesheet structure for raw sequencing reads submission. The input format follows [assets/schema_input_reads.json](../assets/schema_input_reads.json). +In addition to the [common parameters](#running-the-pipeline): -Example: +| Parameter | Required | Description | +| -------------- | -------- | -------------------------------------------------------------------------- | +| `--upload_tpa` | No | Mark assemblies as third party assemblies when required. Default: `false`. | -```csv title="samplesheet_reads.csv" -sample,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,library_selection,library_strategy,insert_size,library_name,description -illumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLUMINA,Illumina HiSeq 2000,GENOMIC,RANDOM,WGS,500,HiSeq_library_001,Illumina sequencing of sample XYZ -pacbio_run_001,SAMEA7654321,data/pacbio_reads.fastq.gz,,PACBIO_SMRT,PacBio Sequel,GENOMIC,RANDOM,WGS,,PacBio_library_002,Long-read sequencing +Example `metagenomic_assemblies` mode upload to TEST server, run command with docker: + +```bash +nextflow run nf-core/seqsubmit \ + -profile docker \ + --mode metagenomic_assemblies \ + --input samplesheet_assembly.csv \ + --submission_study \ + --webincli_mode submit \ + --test_upload \ + --outdir assembly_submission ``` -> [!IMPORTANT] -> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. +Example `metagenomic_assemblies` mode upload to LIVE server, run command with docker: -| Column | Required | Description | -| ------------------- | -------- | --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | Yes | Unique identifier of this particular data entry. Used as an experiment name. | -| `sample_accession` | Yes | ENA sample accession (starting with SAMEA) of the sample used to generate raw reads. | -| `fastq_1` | Yes | Path to forward reads in FASTQ format (optionally gzipped). | -| `fastq_2` | No | Path to reverse reads for paired-end data. Leave empty for single-end reads. | -| `platform` | Yes | Sequencing platform. Supported values: `ILLUMINA`, `PACBIO_SMRT`, `OXFORD_NANOPORE`, `ION_TORRENT`, `CAPILLARY`, `DNBSEQ`, `ELEMENT`, `GENAPSYS`, `GENEMIND`, `HELICOS`, `LS454`, `BGISEQ`, `ULTIMA`, `VELA_DIAGNOSTICS`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | -| `instrument` | Yes | Sequencer model, e.g. "Illumina HiSeq 2000", "PacBio Sequel", "MinION". See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#permitted-values-for-instrument) | -| `library_source` | Yes | Library source type. Options: `GENOMIC`, `METAGENOMIC`, `TRANSCRIPTOMIC`, `METAGENOMIC SINGLE CELL`, `TRANSCRIPTOMIC SINGLE CELL`, `SYNTHETIC`, `VIRAL RNA`, `OTHER`. | -| `library_selection` | Yes | Library selection method. Options: `RANDOM`, `PCR`, `RANDOM PCR`, `RT-PCR`, `MF`, `cDNA`, `cDNA_randomPriming`, `cDNA_oligo_dT`, `PolyA`, `Inverse rRNA`, `ChIP`, `MNase`, `DNase`, `Hybrid Selection`, etc. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | -| `library_strategy` | Yes | Library strategy. Options: `WGS`, `WGA`, `WXS`, `RNA-Seq`, `miRNA-Seq`, `ncRNA-Seq`, `EST`, `Hi-C`, `ATAC-seq`, `WCS`, `RAD-Seq`, `CLONE`, `AMPLICON`, `POOLCLONE`, `etc`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | -| `insert_size` | No | Fragment/insert size for paired-end reads (e.g., 500 for 500 bp inserts). Leave empty if not applicable. | -| `library_name` | No | Descriptive library name (optional). | -| `description` | No | Free-text description of the experiment (optional). | +```bash +nextflow run nf-core/seqsubmit \ + -profile docker \ + --mode metagenomic_assemblies \ + --input samplesheet_assembly.csv \ + --submission_study \ + --test_upload false \ + --webincli_mode submit \ + --outdir assemblies_submission/ +``` -## Data privacy +### Limitations -You can reference private ENA data if you have access to it via your Webin account (it was submitted previously under your credentials). To use private data accessions in your submission metadata, specify the `--is_private` flag. The pipeline will then use your provided Webin credentials to fetch the required metadata. +`metagenomic_assemblies` mode does not yet support the following: -You can also control the privacy of your submitted data: +- **Co-assemblies**: Metagenomic co-assemblies are not supported. The pipeline currently expects each assembly to be derived from a single run. Tracked in [issue #61](https://github.com/nf-core/seqsubmit/issues/61), addressed in [PR #66](https://github.com/nf-core/seqsubmit/pull/66). -- If you provide an existing study via `--submission_study`, your submission will inherit the same privacy status as that study. -- If no study is provided, the pipeline will register a new one for you. To keep this new study private, you must specify a release date using `--release_date YYYY-MM-DD` (up to 2 years from the current date). +- **Submission of an assembly without submitting the raw reads**: Assemblies that have no associated reads accession (`run_accession` column in the samplesheet) are not supported, since the pipeline relies on that lineage to build the required metadata. Tracked in [issue #29](https://github.com/nf-core/seqsubmit/issues/29). + +- **Single-contig assemblies**: Assemblies that consist of only one contig cannot be uploaded through this pipeline. ENA classifies these as "chromosomal assemblies", which follow a different submission procedure with different metadata requirements. If you have a single-contig assembly to submit, please contact [ENA support](https://www.ebi.ac.uk/ena/browser/support) for guidance. + +- **Coverage calculation from long reads**: `coverage` calculation via `coverm` has only been tested and validated with short reads (e.g. Illumina). It has not been tested with long reads (PacBio, Nanopore). Exercise caution when processing data generated from long reads, as issues may occur. + +## `mags` and `bins` modes + +### Prerequisites + +In addition to the general [prerequisites](#before-you-start), for `mags` or `bins` mode: + +- The raw reads only, or raw reads and metagenomic assemblies, used to generate the MAGs/bins have to be submitted to INSDC/ENA to obtain the corresponding run or assembly accessions. Refer to the [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads.html) for details. You can use the pipeline in `reads` and `metagenomic_assemblies` mode to do this. Assigned run or assembly accessions have to be provided in `accession` column of the samplesheet. + +### Samplesheet input + +You will need to create a samplesheet with information about the MAGs/bins you would like to submit before running the pipeline. Use `--input` to specify its location. It must be a comma-separated file with the structure that follows [assets/schema_input_genome.json](../assets/schema_input_genome.json). + +Example: + +```csv title="samplesheet_genomes.csv" +sample,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage +mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default,CheckM2 1.0.1,92.81,1.09,66.04,sediment metagenome,No,marine,cable bacteria,marine sediment,No,d__Bacteria;p__Proteobacteria;s__ +``` + +> [!IMPORTANT] +> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. + +| Column | Required | Description | +| --------------------------- | ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `sample` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset. | +| `fasta` | Yes | Path to MAG/bin contigs in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. | +| `accession` | Yes | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | +| `fastq_1` | Conditional | Path to the read file in FASTQ format used to generate the source metagenomic assembly. Required if `genome_coverage` is not provided. | +| `fastq_2` | No | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | +| `assembly_software` | Yes | Tool name and version that were used to generate the source metagenomic assembly. | +| `binning_software` | Yes | Binning tool, including version, that was used to generate the bins. | +| `binning_parameters` | Yes | Arguments that were used during binning. | +| `stats_generation_software` | No | Tool, including version, that was used to calculate completeness and contamination. | +| `completeness` | No | Genome completeness value. | +| `contamination` | No | Genome contamination value. | +| `genome_coverage` | Conditional | Estimated average sequencing depth across the genome. If the value is missing, it is computed automatically during pipeline execution when reads are provided. | +| `metagenome` | Yes | Registered metagenome taxonomic identifier or name that matches an existing ENA taxonomy entry. For more details see https://ena-docs.readthedocs.io/en/latest/faq/taxonomy.html | +| `co-assembly` | Yes | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: Yes or No. | +| `broad_environment` | Yes | Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428) | +| `local_environment` | Yes | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". | +| `environmental_medium` | Yes | Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). | +| `RNA_presence` | No | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: Yes or No. | +| `NCBI_lineage` | No | NCBI taxonomy lineage of the genome. Can be composted of either numerical IDs or official NCBI taxon names separated by ";". | -| Example | Source Data | Submission Visibility | Required Arguments | Result | -| --------------------------------------------------------------------------------------------------------------------------------------------------------------------- | ----------- | --------------------- | ------------------------------------------------------ | ----------------------------------------------------- | -| Assemblies generated from public reads. Assemblies should be public immediately after submission | public | public | – | SUCCESS | -| Assemblies generated from public reads. Assemblies should remain private for 1 year after submission | public | private | `--release_date YYYY-MM-DD` (+1 year) | SUCCESS | -| Bins generated from private reads and assemblies. Bins should be public after submission. User **has access** to the reads and assemblies via Webin account | private | public | `--is_private` | SUCCESS | -| Bins generated from private reads and assemblies. Bins should remain private for 2 years after submission. User **has access** via Webin account | private | private | `--is_private`, `--release_date YYYY-MM-DD` (+2 years) | SUCCESS | -| MAGs generated from private reads and assemblies. MAGs should be public after submission. User **does not have access** to the reads and assemblies via Webin account | private | public | – | ERROR (submission can only reference accessible data) | +> [!NOTE] +> More information about environment tags can be found at checklists [ERC000050](https://www.ebi.ac.uk/ena/browser/view/ERC000050) for bins and [ERC000047](https://www.ebi.ac.uk/ena/browser/view/ERC000047) for MAGs under the field names "broad-scale environmental context", "local environmental context", and "environmental medium". + +If `genome_coverage`, `stats_generation_software`, `completeness`, `contamination`, `RNA_presence`, or `NCBI_lineage` are missing, the workflow can calculate or infer them when the required inputs are available. See the [Methods documentation](docs/methods.md) for more information on how metadata statistics are obtained. -## Database preparation (`mags` / `bins`) +### Database preparation (`CheckM2` and `CAT_pack`) -The `GENOMESUBMIT` workflow uses `CheckM2` and `CAT_pack` that require specialized databases for completeness/contamination assessment and taxonomy assignment. +`mags` and `bins` submission modes use `CheckM2` and `CAT_pack` tools that require specialized databases for completeness/contamination assessment and taxonomy assignment. You can either provide pre-existing databases or let the pipeline prepare them during execution. @@ -253,75 +336,93 @@ In particular the `CAT_pack` database is extremely large and may take a long tim See [CAT_pack documentation](https://github.com/MGXlab/CAT_pack) and [CheckM2 documentation](https://github.com/chklovski/CheckM2) for more details on usage and creation of databases. Databases created/downloaded by the pipeline are published under: -`/databases/`. In the subsequent pipeline runs it is strongly recommended to reuse them using `--cat_db /databases/cat_pack/` and `--cat_db /databases/checkm2/`. +`/databases/`. In the subsequent pipeline runs you can reuse them using `--cat_db /databases/cat_pack/` and `--cat_db /databases/checkm2/`. -## Running the pipeline +### Running the pipeline in `mags` and `bins` modes -General command template: +In addition to the [common parameters](#running-the-pipeline): -```bash -nextflow run nf-core/seqsubmit \ - -profile \ - --mode \ - --input \ - --centre_name \ - --submission_study \ - --outdir -``` - -Key parameters: +| Parameter | Required | Description | +| -------------- | -------- | ------------------------------------------------------------------------------------------------------------------------------------------------ | +| `--upload_tpa` | No | Mark assemblies as third party assemblies when required. Default: `false`. | +| `--checkm2_db` | No | Path to a local `CheckM2` database. If omitted, downloads the version set by `--checkm2_db_download_id`. Default: `14897628` (Zenodo accession). | +| `--cat_db` | No | Path to a local `CAT_pack` database. If omitted, constructs the version set by `--cat_db_download_id`. Default: `nr`. | -| Parameter | Description | -| -------------------- | ------------------------------------------------------------------------------------------------------------------------------------ | -| `--mode` | Submission type. Supported values are `mags`, `bins`, `metagenomic_assemblies`, and `reads`. | -| `--input` | Path to the samplesheet describing the data to submit. | -| `--submission_study` | ENA study accession (PRJ/ERP) to submit the data to. For metagenomic assemblies, this is the paper's ENA Assembly Project accession. | -| `--centre_name` | Name of the submitter's organisation. | -| `--test_upload` | Submit to the ENA TEST server instead of the LIVE server. | -| `--webincli_mode` | Webin-CLI mode for ENA interaction: `submit` uploads data, `validate` performs validation only. | -| `--upload_tpa` | Mark assemblies as third party assemblies when required. | - -Test example for `mags` run with docker: +Example `mags` mode upload to TEST server, run command with docker: ```bash nextflow run nf-core/seqsubmit \ -profile docker \ --mode mags \ - --input assets/samplesheet_genomes.csv \ + --input samplesheet_genomes.csv \ --submission_study \ - --centre_name TEST_CENTER \ + --centre_name \ --webincli_mode submit \ --test_upload \ - --outdir results/validate_mags + --outdir mags_submission/ ``` -Test example for `metagenomic_assemblies` run with docker: +### Limitations -```bash -nextflow run nf-core/seqsubmit \ - -profile docker \ - --mode metagenomic_assemblies \ - --input assets/samplesheet_assembly.csv \ - --submission_study \ - --centre_name TEST_CENTER \ - --webincli_mode submit \ - --test_upload \ - --outdir results/validate_assemblies -``` +`mags` and `bins` modes do not yet support the following: + +- **Co-assemblies**: MAGs/bins generated from co-assemblies are not supported. The pipeline currently expects each MAG/bin to be derived from an assembly generated from a single run. Tracked in [issue #61](https://github.com/nf-core/seqsubmit/issues/61). + +- **Eukaryotic and viral bins/MAGs**: Submitting eukaryotic or viral genomes that require additional metadata generation is not supported. For such genomes tRNA and rRNA prediction, quality assessment (completeness/contamination estimation) and taxonomy assignment are not implemented. If all the required metadata (`completeness`, `contamination`, `stats_generation_software`, `RNA_presence` and `NCBI_lineage`) is already provided in the samplesheet, eukaryotic and viral bins/MAGs can still be submitted normally. Tracked in [issue #40](https://github.com/nf-core/seqsubmit/issues/40) (eukaryotic) and [issue #63](https://github.com/nf-core/seqsubmit/issues/63) (viral). -Test example for `reads` run with docker: +- **Submission of a MAG/bin without submitting the assembly or raw reads**: If a MAG/bin cannot be traced back to an assembly accession or a reads accession — i.e. only a sample accession is available — it cannot currently be submitted, since the pipeline relies on that lineage to build the required metadata. Tracked in [issue #87](https://github.com/nf-core/seqsubmit/issues/87). + +- **Single-contig MAGs/bins**: MAGs/bins that consist of only one contig cannot be uploaded through this pipeline. ENA classifies these as "chromosomal assemblies", which follow a different submission procedure with different metadata requirements. If you have a single-contig MAG/bin to submit, please contact [ENA support](https://www.ebi.ac.uk/ena/browser/support) for guidance. + +- **Coverage calculation from long reads**: `genome_coverage` calculation via `coverm` has only been tested and validated with short reads (e.g. Illumina). It has not been tested with long reads (PacBio, Nanopore). Exercise caution when processing data generated from long reads, as issues may occur. + +## Data privacy + +You can reference private ENA data if you have access to it via your Webin account (it was submitted previously under your credentials). To use private data accessions in your submission metadata, specify the `--is_private` flag. The pipeline will then use your provided Webin credentials to fetch the required metadata. + +You can also control the privacy of your submitted data: + +- If you provide an existing study via `--submission_study`, your submission will inherit the same privacy status as that study. +- If no study is provided, the pipeline will register a new one for you. To keep this new study private, you must specify a release date using `--release_date YYYY-MM-DD` (up to 2 years from the current date). + +| Example | Source Data | Submission Visibility | Required Arguments | Result | +| --------------------------------------------------------------------------------------------------------------------------------------------------------------------- | ----------- | --------------------- | ---------------------------------------- | ----------------------------------------------------- | +| Assemblies generated from public reads. Assemblies should be public immediately after submission | public | public | – | SUCCESS | +| Assemblies generated from public reads. Assemblies should remain private until 1 July 2028 submission | public | private | `--release_date 2028-07-01` | SUCCESS | +| Bins generated from private reads and assemblies. Bins should be public after submission. User **has access** to the reads and assemblies via Webin account | private | public | `--is_private` | SUCCESS | +| Bins generated from private reads and assemblies. Bins should remain private until 1 July 2028. User **has access** via Webin account | private | private | `--is_private --release_date 2028-07-01` | SUCCESS | +| MAGs generated from private reads and assemblies. MAGs should be public after submission. User **does not have access** to the reads and assemblies via Webin account | private | public | – | ERROR (submission can only reference accessible data) | + +## Running the pipeline + +General command template: ```bash nextflow run nf-core/seqsubmit \ - -profile docker \ - --mode reads \ - --input samplesheet_reads.csv \ + -profile \ + --mode \ + --input \ + --centre_name \ --submission_study \ - --webincli_mode submit \ - --test_upload \ - --outdir results/validate_reads + --outdir ``` +Parameters common to all modes: + +| Parameter | Required | Description | +| ------------------------------------------ | ------------ | -------------------------------------------------------------------------------------------------------------------- | +| `--mode` | Yes | Type of the data to be submitted. Options: `mags`, `bins`, `metagenomic_assemblies`, `reads`. | +| `--input` | Yes | Path to the samplesheet describing the data to submit. | +| `--outdir` | Yes | Path to the output directory for pipeline results. | +| `--submission_study` OR `--study_metadata` | Yes (one of) | ENA study accession (PRJ/ERP) to submit the data to OR metadata file in JSON/TSV/CSV format to register a new study. | +| `--centre_name` | Yes | Name of the submitter's organisation. | +| `--test_upload` | No | Submit to the ENA TEST server instead of the LIVE server. Default: `true`. | +| `--webincli_mode` | No | Webin-CLI mode for ENA interaction: `submit` uploads data, `validate` performs validation only. Default: `submit`. | +| `--is_private` | No | Use if you are referencing private data accessions in your submission. Default: `false`. | +| `--release_date` | No | Use if you want to keep your data private after submission until a particular date. | + +Each mode also has additional parameters and example commands — see its own "Running the pipeline in ... mode" subsection above. + If you wish to repeatedly use the same parameters for multiple runs, rather than specifying each flag in the command, you can specify these in a params file. Pipeline settings can be provided in a `yaml` or `json` file via `-params-file `. From b61853e6d0d4125ab9bed3c8cdbf964dcc4f3500 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Fri, 17 Jul 2026 10:00:41 +0000 Subject: [PATCH 24/47] polish usage.md structure and wording --- docs/usage.md | 115 ++++++++++++++++++++++++-------------------------- 1 file changed, 56 insertions(+), 59 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index aa43cc0..5a1fb8a 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -9,26 +9,12 @@ - [Introduction](#introduction) - [Before you start](#before-you-start) - [Submission study](#submission-study) +- [Data privacy](#data-privacy) - [`reads` mode](#reads-mode) - - [Prerequisites](#prerequisites) - - [Samplesheet input](#samplesheet-input) - - [Running the pipeline in `reads` mode](#running-the-pipeline-in-reads-mode) - - [Limitations](#limitations) - [`metagenomic_assemblies` mode](#metagenomic_assemblies-mode) - - [Prerequisites](#prerequisites-1) - - [Samplesheet input](#samplesheet-input-1) - - [Running the pipeline in `metagenomic_assemblies` mode](#running-the-pipeline-in-metagenomic_assemblies-mode) - - [Limitations](#limitations-1) - [`mags` and `bins` modes](#mags-and-bins-modes) - - [Prerequisites](#prerequisites-2) - - [Samplesheet input](#samplesheet-input-2) - - [Database preparation (`CheckM2` and `CAT_pack`)](#database-preparation-checkm2-and-cat_pack) - - [Running the pipeline in `mags` and `bins` modes](#running-the-pipeline-in-mags-and-bins-modes) - - [Limitations](#limitations-2) -- [Data privacy](#data-privacy) +- [Scalability](#scalability) - [Running the pipeline](#running-the-pipeline) - - [Updating the pipeline](#updating-the-pipeline) - - [Reproducibility](#reproducibility) - [Core Nextflow arguments](#core-nextflow-arguments) - [Custom configuration](#custom-configuration) - [Running in the background](#running-in-the-background) @@ -38,13 +24,14 @@ `nf-core/seqsubmit` is a Nextflow pipeline for submitting sequence data to ENA. -The pipeline supports of the following data types, selected via `--mode` parameter: +The pipeline supports the following data types, selected via the `--mode` parameter: - `reads` — raw sequencing reads - `metagenomic_assemblies` — metagenomic assemblies -- `mags` / `bins` — metagenome-assembled genomes (MAGs) or metagenomic bins +- `mags` — metagenome-assembled genomes (MAGs) +- `bins` — metagenomic bins -Each mode has its own samplesheet structure, prerequisites, and limitations — see the dedicated section for the mode you want to use below. The [Before you start](#before-you-start), [Submission study](#submission-study), [Running the pipeline](#running-the-pipeline), and [Data privacy](#data-privacy) sections apply to all modes. +Each mode has its own samplesheet structure, prerequisites, and limitations — see the dedicated section for the mode you want to use below. ## Before you start @@ -60,9 +47,11 @@ Before running the pipeline in any mode, make sure that: nextflow secrets set ENA_WEBIN_PASSWORD "XXX" ``` -- You prepared either study accession or study metadata file to set a submission study to assosiate the submission with. See [Submission study](#submission-study) section below for details. + Make sure to replace the values above with your own credentials. + +- Provide either a study accession or a study registration metadata file for the study the submission will be associated with. See the [Submission study](#submission-study) section below for details. -Each mode also has additional prerequisites — see its own "Prerequisites" subsection below. +- Depending on the chosen mode, samples, reads, or metagenomic assemblies must be pre-submitted to ENA to obtain the corresponding accessions, which you then reference in your submission. Refer to the relevant mode section below for details. ## Submission study @@ -126,13 +115,30 @@ study-soil-2026 Soil microbiome study Survey of soil microbiota | `existing_study_type` | No | ENA study type (e.g. `Metagenomics`, `Other`). | | `new_study_type` | No | Custom study type. Only used when `existing_study_type` is set to `Other`. | +## Data privacy + +You can reference private ENA data if you have access to it via your Webin account (it was submitted previously under your credentials). To use private data accessions in your submission metadata, specify the `--is_private` flag. The pipeline will then use your provided Webin credentials to fetch the required metadata. + +You can also control the privacy of your submitted data: + +- If you provide an existing study via `--submission_study`, your submission will inherit the same privacy status as that study. +- If no study is provided, the pipeline will register a new one for you. To keep this new study private, you must specify a release date using `--release_date YYYY-MM-DD` (up to 2 years from the current date). + +| Example | Source Data | Submission Visibility | Required Arguments | Result | +| --------------------------------------------------------------------------------------------------------------------------------------------------------------------- | ----------- | --------------------- | ---------------------------------------- | ----------------------------------------------------- | +| Assemblies generated from public reads. Assemblies should be public immediately after submission | public | public | – | SUCCESS | +| Assemblies generated from public reads. Assemblies should remain private until 1 July 2028 submission | public | private | `--release_date 2028-07-01` | SUCCESS | +| Bins generated from private reads and assemblies. Bins should be public after submission. User **has access** to the reads and assemblies via Webin account | private | public | `--is_private` | SUCCESS | +| Bins generated from private reads and assemblies. Bins should remain private until 1 July 2028. User **has access** via Webin account | private | private | `--is_private --release_date 2028-07-01` | SUCCESS | +| MAGs generated from private reads and assemblies. MAGs should be public after submission. User **does not have access** to the reads and assemblies via Webin account | private | public | – | ERROR (submission can only reference accessible data) | + ## `reads` mode ### Prerequisites In addition to the general [prerequisites](#before-you-start), for `reads` mode: -- You need to register the source BioSamples and obtain the sample accessions as described in [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/samples.html). Those have to be provided in `sample_accession` column of the samplesheet. +- You need to register the source BioSamples and obtain the sample accessions as described in the [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/samples.html). These accessions must be provided in the `sample_accession` column of the samplesheet. ### Samplesheet input @@ -164,7 +170,7 @@ pacbio_run_001,SAMEA7654321,data/pacbio_reads.fastq.gz,,PACBIO_SMRT,PacBio Seque | `library_name` | No | Descriptive library name (optional). | | `description` | No | Free-text description of the experiment (optional). | -### Running the pipeline in `reads` mode +### `reads` parameters and example `reads` mode does not add any parameters beyond the [common parameters](#running-the-pipeline). @@ -193,7 +199,7 @@ nextflow run nf-core/seqsubmit \ In addition to the general [prerequisites](#before-you-start), for `metagenomic_assemblies` mode: -- The raw reads used to generate the assemblies have have to be submitted to INSDC/ENA to obtain the corresponding run accessions. Refer to the [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads.html) for details. You can use `seqsubmit` in `reads` mode to do this. Assigned run accessions have to be provided in `run_accession` column of the samplesheet. +- The raw reads used to generate the assemblies have to be submitted to INSDC/ENA to obtain the corresponding run accessions. Refer to the [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads.html) for details. You can use `seqsubmit` in `reads` mode to do this. Assigned run accessions must be provided in the `run_accession` column of the samplesheet. ### Samplesheet input @@ -221,9 +227,9 @@ assembly_002,data/assembly_002.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 | `assembler` | Yes | Name of the assembler software used to generate the assembly. | | `assembler_version` | Yes | Version of the assembler software used to generate the assembly. | -Provide either read files (`fastq_1`, optionally `fastq_2`) or a `coverage` value for each row. If `coverage` is missing and reads are provided, the workflow calculates average coverage with `coverm`. See the [Methods documentation](docs/methods.md) for more information on how coverage is calculated. +Provide either read files (`fastq_1`, optionally `fastq_2`) or a `coverage` value for each row. If `coverage` is missing and reads are provided, the workflow calculates average coverage with CoverM. See the [Methods documentation](docs/methods.md) for more information on how coverage is calculated. -### Running the pipeline in `metagenomic_assemblies` mode +### `metagenomic_assemblies` parameters and example In addition to the [common parameters](#running-the-pipeline): @@ -267,7 +273,7 @@ nextflow run nf-core/seqsubmit \ - **Single-contig assemblies**: Assemblies that consist of only one contig cannot be uploaded through this pipeline. ENA classifies these as "chromosomal assemblies", which follow a different submission procedure with different metadata requirements. If you have a single-contig assembly to submit, please contact [ENA support](https://www.ebi.ac.uk/ena/browser/support) for guidance. -- **Coverage calculation from long reads**: `coverage` calculation via `coverm` has only been tested and validated with short reads (e.g. Illumina). It has not been tested with long reads (PacBio, Nanopore). Exercise caution when processing data generated from long reads, as issues may occur. +- **Coverage calculation from long reads**: `coverage` calculation via CoverM has only been tested and validated with short reads (e.g. Illumina). It has not been tested with long reads (PacBio, Nanopore). Exercise caution when processing data generated from long reads, as issues may occur. ## `mags` and `bins` modes @@ -275,7 +281,7 @@ nextflow run nf-core/seqsubmit \ In addition to the general [prerequisites](#before-you-start), for `mags` or `bins` mode: -- The raw reads only, or raw reads and metagenomic assemblies, used to generate the MAGs/bins have to be submitted to INSDC/ENA to obtain the corresponding run or assembly accessions. Refer to the [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads.html) for details. You can use the pipeline in `reads` and `metagenomic_assemblies` mode to do this. Assigned run or assembly accessions have to be provided in `accession` column of the samplesheet. +- Either the raw reads alone, or the raw reads together with the metagenomic assemblies, used to generate the MAGs/bins must be submitted to INSDC/ENA to obtain the corresponding run or assembly accessions. Refer to the [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads.html) for details. You can use the pipeline in `reads` and `metagenomic_assemblies` mode to do this. Assigned run or assembly accessions must be provided in the `accession` column of the samplesheet. ### Samplesheet input @@ -318,35 +324,35 @@ mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default, If `genome_coverage`, `stats_generation_software`, `completeness`, `contamination`, `RNA_presence`, or `NCBI_lineage` are missing, the workflow can calculate or infer them when the required inputs are available. See the [Methods documentation](docs/methods.md) for more information on how metadata statistics are obtained. -### Database preparation (`CheckM2` and `CAT_pack`) +### Preparing the CheckM2 and CAT_pack databases -`mags` and `bins` submission modes use `CheckM2` and `CAT_pack` tools that require specialized databases for completeness/contamination assessment and taxonomy assignment. +`mags` and `bins` submission modes use CheckM2 and CAT_pack tools that require specialized databases for completeness/contamination assessment and taxonomy assignment. You can either provide pre-existing databases or let the pipeline prepare them during execution. -As databases preparation can take significant time, we strongly recommend downloading them locally and storing them in a local cache folder for reuse across runs. -In particular the `CAT_pack` database is extremely large and may take a long time to download and compute. +As database preparation can take significant time, we strongly recommend downloading them locally and storing them in a local cache folder for reuse across runs. +In particular, the CAT_pack database is extremely large and may take a long time to download and compute. -- `CheckM2`: - - provide the path to local database with `--checkm2_db`, otherwise the pipeline downloads version specified with `--checkm2_db_download_id` (by default Zenodo accession `14897628`). +- CheckM2: + - provide the path to a local database with `--checkm2_db`, otherwise the pipeline downloads the version specified with `--checkm2_db_download_id` (by default Zenodo accession `14897628`). -- `CAT_pack`: - - provide the path to local database (containing `tax/` and `db/` folders or `tar.gz` archive) with `--cat_db`, otherwise the pipeline constructs version specified with `--cat_db_download_id` (by default `nr`). +- CAT_pack: + - provide the path to a local database (containing `tax/` and `db/` folders or a `tar.gz` archive) with `--cat_db`, otherwise the pipeline constructs the version specified with `--cat_db_download_id` (by default `nr`). -See [CAT_pack documentation](https://github.com/MGXlab/CAT_pack) and [CheckM2 documentation](https://github.com/chklovski/CheckM2) for more details on usage and creation of databases. +See the [CAT_pack documentation](https://github.com/MGXlab/CAT_pack) and [CheckM2 documentation](https://github.com/chklovski/CheckM2) for more details on usage and creation of databases. Databases created/downloaded by the pipeline are published under: -`/databases/`. In the subsequent pipeline runs you can reuse them using `--cat_db /databases/cat_pack/` and `--cat_db /databases/checkm2/`. +`/databases/`. In subsequent pipeline runs you can reuse them using `--cat_db /databases/cat_pack/` and `--checkm2_db /databases/checkm2/`. -### Running the pipeline in `mags` and `bins` modes +### `mags` and `bins` parameters and example In addition to the [common parameters](#running-the-pipeline): -| Parameter | Required | Description | -| -------------- | -------- | ------------------------------------------------------------------------------------------------------------------------------------------------ | -| `--upload_tpa` | No | Mark assemblies as third party assemblies when required. Default: `false`. | -| `--checkm2_db` | No | Path to a local `CheckM2` database. If omitted, downloads the version set by `--checkm2_db_download_id`. Default: `14897628` (Zenodo accession). | -| `--cat_db` | No | Path to a local `CAT_pack` database. If omitted, constructs the version set by `--cat_db_download_id`. Default: `nr`. | +| Parameter | Required | Description | +| -------------- | -------- | ---------------------------------------------------------------------------------------------------------------------------------------------- | +| `--upload_tpa` | No | Mark assemblies as third party assemblies when required. Default: `false`. | +| `--checkm2_db` | No | Path to a local CheckM2 database. If omitted, downloads the version set by `--checkm2_db_download_id`. Default: `14897628` (Zenodo accession). | +| `--cat_db` | No | Path to a local CAT_pack database. If omitted, constructs the version set by `--cat_db_download_id`. Default: `nr`. | Example `mags` mode upload to TEST server, run command with docker: @@ -374,24 +380,15 @@ nextflow run nf-core/seqsubmit \ - **Single-contig MAGs/bins**: MAGs/bins that consist of only one contig cannot be uploaded through this pipeline. ENA classifies these as "chromosomal assemblies", which follow a different submission procedure with different metadata requirements. If you have a single-contig MAG/bin to submit, please contact [ENA support](https://www.ebi.ac.uk/ena/browser/support) for guidance. -- **Coverage calculation from long reads**: `genome_coverage` calculation via `coverm` has only been tested and validated with short reads (e.g. Illumina). It has not been tested with long reads (PacBio, Nanopore). Exercise caution when processing data generated from long reads, as issues may occur. - -## Data privacy +- **Coverage calculation from long reads**: `genome_coverage` calculation via CoverM has only been tested and validated with short reads (e.g. Illumina). It has not been tested with long reads (PacBio, Nanopore). Exercise caution when processing data generated from long reads, as issues may occur. -You can reference private ENA data if you have access to it via your Webin account (it was submitted previously under your credentials). To use private data accessions in your submission metadata, specify the `--is_private` flag. The pipeline will then use your provided Webin credentials to fetch the required metadata. +## Scalability -You can also control the privacy of your submitted data: +Some processes in this pipeline are resource intensive — in particular coverage calculation (CoverM), quality assessment (CheckM2), and taxonomic classification of MAGs/bins (BAT). For real-world data, these steps should be run on an HPC cluster or another compute environment with sufficient CPU, memory, and job scheduling capacity, rather than on a single local machine. -- If you provide an existing study via `--submission_study`, your submission will inherit the same privacy status as that study. -- If no study is provided, the pipeline will register a new one for you. To keep this new study private, you must specify a release date using `--release_date YYYY-MM-DD` (up to 2 years from the current date). +Depending on the resources available on your machine or cluster, you will likely need to adapt the Nextflow configuration to limit the maximum number of concurrently running jobs and other resource settings (e.g. `max_cpus`, `max_memory`, `max_time`, or `executor.queueSize` in a custom config), so that the pipeline does not try to claim more resources than are actually available. See the [nf-core configuration documentation](https://nf-co.re/docs/usage/getting_started/configuration) for guidance on writing a custom config for your infrastructure. -| Example | Source Data | Submission Visibility | Required Arguments | Result | -| --------------------------------------------------------------------------------------------------------------------------------------------------------------------- | ----------- | --------------------- | ---------------------------------------- | ----------------------------------------------------- | -| Assemblies generated from public reads. Assemblies should be public immediately after submission | public | public | – | SUCCESS | -| Assemblies generated from public reads. Assemblies should remain private until 1 July 2028 submission | public | private | `--release_date 2028-07-01` | SUCCESS | -| Bins generated from private reads and assemblies. Bins should be public after submission. User **has access** to the reads and assemblies via Webin account | private | public | `--is_private` | SUCCESS | -| Bins generated from private reads and assemblies. Bins should remain private until 1 July 2028. User **has access** via Webin account | private | private | `--is_private --release_date 2028-07-01` | SUCCESS | -| MAGs generated from private reads and assemblies. MAGs should be public after submission. User **does not have access** to the reads and assemblies via Webin account | private | public | – | ERROR (submission can only reference accessible data) | +When submitting a large number of records, be aware that ENA recommends limiting submissions to 5000 per day to avoid overfilling their processing queue. If you need to submit more than this, consider splitting your samplesheet and spreading the submission across several days. ## Running the pipeline @@ -421,7 +418,7 @@ Parameters common to all modes: | `--is_private` | No | Use if you are referencing private data accessions in your submission. Default: `false`. | | `--release_date` | No | Use if you want to keep your data private after submission until a particular date. | -Each mode also has additional parameters and example commands — see its own "Running the pipeline in ... mode" subsection above. +Each mode also has additional parameters and example commands — see its own "`` parameters and example" subsection above. If you wish to repeatedly use the same parameters for multiple runs, rather than specifying each flag in the command, you can specify these in a params file. 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zRCP%tyi)wjOK80Do?kxOTbAB-VXHXxtCigB9p1y!B-glL8XM|7D;#mXbV)!(MMd$h z5hDuUuy(nh?yqD9^Ho`qS8MN{LAf+e`(sH{2SIXQEXI-}emyc9Sl>Xq?7QfVLGg;; zIpujmMeg0kATi{@4#6DXYWN?@vK%2N~41-U@7FL!UJ5n3>dsotIj0&tNog1P}EE zVfKs7t56--WmdJ$vW|&neSVcX>Z;e+n2~lzRjaX8_;Qj> z)KOHFLewI-Zs)r=&w}c313`yYtV6BCqQwIH`ubdt9i@J~P+w~sl8nS*y!(n9{W0h{ zA|26RN`a`rcpc`|kr)eo{9j{=|4rc~fZ - - - - - - - - - - - - - + + + + + + + + + + + + + From 2db92bd294c3255d8f53860e992afbe927216d62 Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Fri, 17 Jul 2026 10:20:12 +0000 Subject: [PATCH 26/47] restructure README --- README.md | 208 ++++++++++++------------------------------------------ 1 file changed, 44 insertions(+), 164 deletions(-) diff --git a/README.md b/README.md index e66d1de..3059b1e 100644 --- a/README.md +++ b/README.md @@ -19,70 +19,56 @@ [![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23seqsubmit-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/seqsubmit)[![Follow on Bluesky](https://img.shields.io/badge/bluesky-%40nf__core-1185fe?labelColor=000000&logo=bluesky)](https://bsky.app/profile/nf-co.re)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core) -## Table of Contents - -- [Introduction](#introduction) -- [Requirements](#requirements) -- [Input samplesheets](#input-samplesheets) -- [Usage](#usage) - - [Submission study](#submission-study) - - [Data privacy](#data-privacy) - - [Database setup (`CheckM2` and `CAT_pack`)](#database-setup-checkm2-and-cat_pack) - - [Required parameters:](#required-parameters) - - [Optional parameters:](#optional-parameters) -- [Pipeline output](#pipeline-output) -- [Scalability](#scalability) -- [Limitations](#limitations) -- [Credits](#credits) -- [Contributions and Support](#contributions-and-support) -- [Citations](#citations) - ## Introduction +![seqsubmit workflow diagram](assets/seqsubmit_schema.png) + **nf-core/seqsubmit** is a Nextflow pipeline for submitting sequence data to [ENA](https://www.ebi.ac.uk/ena/browser/home). -Currently, the pipeline supports four submission modes, each routed to a dedicated workflow and requiring its own input samplesheet structure: +The pipeline currently supports the following submission modes, each routed to a dedicated workflow: -- `mags` for Metagenome Assembled Genomes (MAGs) submission with `GENOMESUBMIT` workflow -- `bins` for bins submission with `GENOMESUBMIT` workflow -- `metagenomic_assemblies` for assembly submission with `ASSEMBLYSUBMIT` workflow -- `reads` for raw sequencing reads submission with `READSUBMIT` workflow +- `reads` — raw sequencing reads submission via the `READSUBMIT` workflow (pink) +- `metagenomic_assemblies` — assembly submission via the `ASSEMBLYSUBMIT` workflow (green) +- `mags` — metagenome-assembled genomes (MAGs) submission via the `GENOMESUBMIT` workflow (blue) +- `bins` — bins submission via the `GENOMESUBMIT` workflow (blue) -![seqsubmit workflow diagram](assets/seqsubmit_schema.png) + + +Each workflow has its own samplesheet structure, prerequisites, and limitations — they are briefly described below. See the [usage documentation](https://nf-co.re/seqsubmit/usage) for more detailed explanations. ## Requirements - [Nextflow](https://www.nextflow.io/) `>=25.04.0` -- Webin account registered at https://www.ebi.ac.uk/ena/submit/webin/login -- For metagenomic assemblies submission the raw reads used to generate the assemblies have to be submitted to INSDC/ENA to obtain the corresponding run accessions. You can use the pipeline in mode `reads` to perform this. -- For MAGs or bins submission the raw reads only or raw reads and metagenomic assemblies used to generate the MAGs/bins have to be submitted to INSDC/ENA to obtain the corresponding run or assembly accessions. You can use the pipeline in mode `reads` or `metagenomic_assemblies` to perform this. -- For raw reads submission it is required to register source BioSamples and obtain the sample accessions. +- A Webin account registered at -Setup Webin credentials as Nextflow secrets before running the pipeline: + Set your Webin credentials as Nextflow secrets: -`nextflow secrets set ENA_WEBIN "Webin-XXX"` + ```bash + nextflow secrets set ENA_WEBIN "Webin-XXX" + nextflow secrets set ENA_WEBIN_PASSWORD "XXX" + ``` -`nextflow secrets set ENA_WEBIN_PASSWORD "XXX"` + Make sure to replace the values above with your own credentials. -Make sure you update commands above with your authorised credentials. +- Provide either a study accession or a study registration metadata file for the study the submission will be associated with. See the [Submission study](docs/usage.md#submission-study) section of the usage documentation for details. -## Input samplesheets +- Depending on the chosen mode, samples, reads, or metagenomic assemblies must be pre-submitted to ENA to obtain the corresponding accessions, which you then reference in your submission. Refer to the relevant mode section in the [usage documentation](https://nf-co.re/seqsubmit/usage) for details. -### `mags` and `bins` modes +## Input samplesheets -The input must follow [assets/schema_input_genome.json](assets/schema_input_genome.json). See the [corresponding samplesheet section](docs/usage.md#mags-and-bins-modes-genomesubmit) of the usage documentation for the full list of required and optional columns. +### `reads` mode -Example `samplesheet_genomes.csv`: +Example: ```csv -sample,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage -lachnospira_eligens,data/bin_lachnospira_eligens.fa.gz,SRR24458089,,,spades_v3.15.5,metabat2_v2.6,default,CheckM2_v1.0.1,61.0,0.21,32.07,sediment metagenome,No,marine,cable_bacteria,marine_sediment,No,d__Bacteria;p__Proteobacteria;s__unclassified_Proteobacteria +sample,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,library_selection,library_strategy,insert_size,library_name,description +illumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLUMINA,Illumina HiSeq 2000,GENOMIC,RANDOM,WGS,500,HiSeq_library_001,Illumina sequencing of sample XYZ ``` -### `metagenomic_assemblies` mode +See the [`reads` mode section](docs/usage.md#samplesheet-input) of the usage documentation for more details. -The input must follow [assets/schema_input_assembly.json](assets/schema_input_assembly.json). See the [corresponding samplesheet section](docs/usage.md#metagenomic_assemblies-mode-assemblysubmit) of the usage documentation for more details. +### `metagenomic_assemblies` mode -Example `samplesheet_assembly.csv`: +Example: ```csv sample,fasta,fastq_1,fastq_2,coverage,run_accession,assembler,assembler_version @@ -90,105 +76,36 @@ assembly_1,data/contigs_1.fasta.gz,data/reads_1.fastq.gz,data/reads_2.fastq.gz,, assembly_2,data/contigs_2.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 ``` -### `reads` mode +See the [`metagenomic_assemblies` mode section](docs/usage.md#samplesheet-input-1) of the usage documentation for more details. -The input must follow [assets/schema_input_reads.json](assets/schema_input_reads.json). See the [corresponding samplesheet section](docs/usage.md#reads-mode-readsubmit) of the usage documentation for more details. +### `mags` and `bins` modes -Example `samplesheet_reads.csv`: +Example: ```csv -sample,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,library_selection,library_strategy,insert_size,library_name,description -illumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLUMINA,Illumina HiSeq 2000,GENOMIC,RANDOM,WGS,500,HiSeq_library_001,Illumina sequencing of sample XYZ +sample,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage +lachnospira_eligens,data/bin_lachnospira_eligens.fa.gz,SRR24458089,,,spades_v3.15.5,metabat2_v2.6,default,CheckM2_v1.0.1,61.0,0.21,32.07,sediment metagenome,No,marine,cable_bacteria,marine_sediment,No,d__Bacteria;p__Proteobacteria;s__unclassified_Proteobacteria ``` +See the [`mags` and `bins` modes section](docs/usage.md#samplesheet-input-2) of the usage documentation for the full list of required and optional columns. + ## Usage > [!NOTE] > If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/get_started/environment_setup/overview) on how to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/get_started/run-your-first-pipeline) with `-profile test` before running the workflow on actual data. -### Submission study - -All data submitted through this pipeline must be associated with an ENA study (project). You can either pass an accession of your existing study via `--submission_study`or provide a metadata file via `--study_metadata` and the pipeline will register the study with ENA before submitting your data. See the [usage documentation](docs/usage.md#submission-study) for more details. - -### Data privacy - -The pipeline supports private submissions. See the [usage documentation](docs/usage.md#data-privacy) for more details. - -### Database setup (`CheckM2` and `CAT_pack`) +### Running the pipeline -The `mags`/`bins` workflow requires databases for completeness/contamination estimation and taxonomy assignment. See [Usage documentation](usage.md) for more details. - -### Required parameters: - -| Parameter | Description | -| ------------------------------------------ | ----------------------------------------------------------------------------------------------------------------- | -| `--mode` | Type of the data to be submitted. Options: `[mags, bins, metagenomic_assemblies, reads]` | -| `--input` | Path to the samplesheet describing the data to be submitted | -| `--outdir` | Path to the output directory for pipeline results | -| `--submission_study` OR `--study_metadata` | ENA study accession (PRJ/ERP) to submit the data to OR metadata file in JSON/TSV/CSV format to register new study | -| `--centre_name` | Name of the submitter's organisation | - -### Optional parameters: - -| Parameter | Description | -| ----------------- | ------------------------------------------------------------------------------------------ | -| `--upload_tpa` | Flag to control the type of assembly study (third party assembly or not). Default: false | -| `--test_upload` | Upload to TEST ENA server instead of LIVE. Default: true | -| `--webincli_mode` | Choose Webin-CLI mode: `submit` or `validate`. Default: `submit` | -| `--is_private` | Use that flag if you are referring private data accessions in your submission | -| `--release_date` | Use that flag if you want to keep your data private after submission until particular date | - -General command template: +Each mode also has its own additional parameters and example commands — see the [usage documentation](https://nf-co.re/seqsubmit/usage) for details. General command template: ```bash nextflow run nf-core/seqsubmit \ - -profile \ - --mode \ - --input \ - --centre_name \ - --submission_study \ - --outdir -``` - -Test run (submission to the ENA TEST server) in `mags` mode: - -```bash -nextflow run nf-core/seqsubmit \ - -profile docker \ - --mode mags \ - --input assets/samplesheet_genomes.csv \ - --submission_study \ - --centre_name TEST_CENTER \ - --webincli_mode submit \ - --test_upload true \ - --outdir results/validate_mags -``` - -Test run (submission to the ENA TEST server) in `metagenomic_assemblies` mode: - -```bash -nextflow run nf-core/seqsubmit \ - -profile docker \ - --mode metagenomic_assemblies \ - --input assets/samplesheet_assembly.csv \ - --submission_study \ - --centre_name TEST_CENTER \ - --webincli_mode submit \ - --test_upload true \ - --outdir results/validate_assemblies -``` - -Live submission example: - -```bash -nextflow run nf-core/seqsubmit \ - -profile docker \ - --mode metagenomic_assemblies \ - --input assets/samplesheet_assembly.csv \ - --submission_study PRJEB98843 \ - --test_upload false \ - --webincli_mode submit \ - --outdir results/live_assembly + -profile \ + --mode \ + --input \ + --centre_name \ + --submission_study \ + --outdir ``` > [!WARNING] @@ -200,51 +117,14 @@ For more details and further functionality, please refer to the [usage documenta Key output locations in `--outdir`: -- `mags/` or `bins/`: genome metadata, manifests, and per-sample submission support files +- `reads/`: per-sample submission receipts and accessions - `metagenomic_assemblies/`: assembly metadata CSVs and per-sample coverage files +- `mags/` or `bins/`: genome metadata, manifests, and per-sample submission support files - `multiqc/`: MultiQC summary report - `pipeline_info/`: execution reports, trace, DAG, and software versions For full details, see the [output documentation](https://nf-co.re/seqsubmit/output). -## Scalability - -Some processes in this pipeline are resource intensive — in particular coverage calculation (`coverm`), quality assessment (`CheckM2`), and taxonomic classification of MAGs/bins (`BAT`). For real life data these steps should be run on an HPC cluster or another compute environment with sufficient CPU, memory, and job scheduling capacity, rather than on a single local machine. - -Depending on the resources available on your machine or cluster, you will likely need to adapt the Nextflow configuration to limit the maximum number of concurrently running jobs and other resource settings (e.g. `max_cpus`, `max_memory`, `max_time`, or `executor.queueSize` in a custom config), so the pipeline doesn't try to claim more resources than are actually available. See the [nf-core configuration documentation](https://nf-co.re/docs/usage/getting_started/configuration) for guidance on writing a custom config for your infrastructure. - -When submitting a large number of records, be aware that ENA recommends limiting submissions to 5000 per day to avoid overfilling their processing queue. If you need to submit more than this, consider splitting your samplesheet and spreading the submission across several days. - -## Limitations - -nf-core/seqsubmit does not yet support the following data types and scenarios, grouped by the mode they affect: - -#### `mags` and `bins` modes (`GENOMESUBMIT`) - -- **Co-assemblies**: MAGs/bins generated from co-assemblies are not supported. The pipeline currently expects each MAG/bin to be derived from an assembly generated from a single run. Tracked in [issue #61](https://github.com/nf-core/seqsubmit/issues/61). - -- **Eukaryotic and viral bins/MAGs**: Submitting eukaryotic or viral genomes that require additional metadata generation is not supported. For such genomes tRNA and rRNA prediction, quality assessment (completeness/contamination estimation) and taxonomy assignment are not implemented. If all the required metadata (`completeness`, `contamination`, `stats_generation_software`, `RNA_presence` and `NCBI_lineage`) is already provided in the samplesheet, eukaryotic and viral bins/MAGs can still be submitted normally. Tracked in [issue #40](https://github.com/nf-core/seqsubmit/issues/40) (eukaryotic) and [issue #63](https://github.com/nf-core/seqsubmit/issues/63) (viral). - -- **Submission of a MAG/bin without submitting the assembly or raw reads**: If a MAG/bin cannot be traced back to an assembly accession or a reads accession — i.e. only a sample accession is available — it cannot currently be submitted, since the pipeline relies on that lineage to build the required metadata. Tracked in [issue #87](https://github.com/nf-core/seqsubmit/issues/87). - -- **Single-contig MAGs/bins**: MAGs/bins that consist of only one contig cannot be uploaded through this pipeline. ENA classifies these as "chromosomal assemblies", which follow a different submission procedure with different metadata requirements. If you have a single-contig MAG/bin to submit, please contact [ENA support](https://www.ebi.ac.uk/ena/browser/support) for guidance. - -- **Coverage calculation from long reads**: `genome_coverage` calculation via `coverm` has only been tested and validated with short reads (e.g. Illumina). It has not been tested with long reads (PacBio, Nanopore). Exercise caution when processing data generated from long reads, as issues may occur. - -#### `metagenomic_assemblies` mode (`ASSEMBLYSUBMIT`) - -- **Co-assemblies**: Metagenomic co-assemblies are not supported. The pipeline currently expects each assembly to be derived from a single run. Tracked in [issue #61](https://github.com/nf-core/seqsubmit/issues/61), addressed in [PR #66](https://github.com/nf-core/seqsubmit/pull/66). - -- **Submission of an assembly without submitting the raw reads**: Assemblies that have no associated reads accession (`run_accession` column in the samplesheet) are not supported, since the pipeline relies on that lineage to build the required metadata. Tracked in [issue #29](https://github.com/nf-core/seqsubmit/issues/29). - -- **Single-contig assemblies**: Assemblies that consist of only one contig cannot be uploaded through this pipeline. ENA classifies these as "chromosomal assemblies", which follow a different submission procedure with different metadata requirements. If you have a single-contig assembly to submit, please contact [ENA support](https://www.ebi.ac.uk/ena/browser/support) for guidance. - -- **Coverage calculation from long reads**: `coverage` calculation via `coverm` has only been tested and validated with short reads (e.g. Illumina). It has not been tested with long reads (PacBio, Nanopore). Exercise caution when processing data generated from long reads, as issues may occur. - -#### `reads` mode (`READSUBMIT`) - -- **Reads in non-FASTQ formats**: Only reads provided as FASTQ (`.fastq.gz`) can be submitted. Other formats (e.g. BAM) are not supported as input. - ## Credits nf-core/seqsubmit was originally written by [Martin Beracochea](https://github.com/mberacochea), [Ekaterina Sakharova](https://github.com/KateSakharova), [Sofia Ochkalova](https://github.com/ochkalova), [Evangelos Karatzas](https://github.com/vagkaratzas) and [Tim Rozday](https://github.com/timrozday-mgnify). From 018d328668b0e10dc1ac2d9e354fef483a43ccbf Mon Sep 17 00:00:00 2001 From: Sofia Ochkalova Date: Fri, 17 Jul 2026 10:51:01 +0000 Subject: [PATCH 27/47] Add section for Third Party Assemblies (TPA) in usage.md --- docs/usage.md | 17 +++++++++++++---- 1 file changed, 13 insertions(+), 4 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index 5a1fb8a..20d377a 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -10,6 +10,7 @@ - [Before you start](#before-you-start) - [Submission study](#submission-study) - [Data privacy](#data-privacy) +- [Third party assemblies (TPA)](#third-party-assemblies-tpa) - [`reads` mode](#reads-mode) - [`metagenomic_assemblies` mode](#metagenomic_assemblies-mode) - [`mags` and `bins` modes](#mags-and-bins-modes) @@ -132,6 +133,14 @@ You can also control the privacy of your submitted data: | Bins generated from private reads and assemblies. Bins should remain private until 1 July 2028. User **has access** via Webin account | private | private | `--is_private --release_date 2028-07-01` | SUCCESS | | MAGs generated from private reads and assemblies. MAGs should be public after submission. User **does not have access** to the reads and assemblies via Webin account | private | public | – | ERROR (submission can only reference accessible data) | +## Third party assemblies (TPA) + +A Third PArty (TPA) Assembly record is an assembly or reassembly built from primary sequence data that has already been submitted to an INSDC database (ENA, GenBank, or DDBJ), rather than newly generated sequence data. TPA records must be built entirely from publicly available accessions or public sequencing reads — they cannot incorporate proprietary or unpublished data, and any newly generated sequence they contain must first be released as a primary submission in its own right. + +If you use `metagenomic_assemblies`, `mags`, or `bins` mode to submit data assembled from public data you did not generate yourself, use `--upload_tpa` to flag the submission as a TPA. + +See [ENA's data policies](https://www.ebi.ac.uk/ena/browser/about/policies) for further guidance on when a TPA designation applies to your submission. + ## `reads` mode ### Prerequisites @@ -233,9 +242,9 @@ Provide either read files (`fastq_1`, optionally `fastq_2`) or a `coverage` valu In addition to the [common parameters](#running-the-pipeline): -| Parameter | Required | Description | -| -------------- | -------- | -------------------------------------------------------------------------- | -| `--upload_tpa` | No | Mark assemblies as third party assemblies when required. Default: `false`. | +| Parameter | Required | Description | +| -------------- | -------- | --------------------------------------------------------------------------------------------------------------- | +| `--upload_tpa` | No | Mark assemblies as [third party assemblies (TPA)](#third-party-assemblies-tpa) when required. Default: `false`. | Example `metagenomic_assemblies` mode upload to TEST server, run command with docker: @@ -350,7 +359,7 @@ In addition to the [common parameters](#running-the-pipeline): | Parameter | Required | Description | | -------------- | -------- | ---------------------------------------------------------------------------------------------------------------------------------------------- | -| `--upload_tpa` | No | Mark assemblies as third party assemblies when required. Default: `false`. | +| `--upload_tpa` | No | Mark assemblies as [third party assemblies (TPA)](#third-party-assemblies-tpa) when required. Default: `false`. | | `--checkm2_db` | No | Path to a local CheckM2 database. If omitted, downloads the version set by `--checkm2_db_download_id`. Default: `14897628` (Zenodo accession). | | `--cat_db` | No | Path to a local CAT_pack database. If omitted, constructs the version set by `--cat_db_download_id`. Default: `nr`. | From 36700b43d30a4c9ec1967f5b52067c9a6af28439 Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Tue, 21 Jul 2026 16:55:57 +0100 Subject: [PATCH 28/47] many fixes --- README.md | 9 +- assets/schema_input_assembly.json | 4 +- assets/schema_input_genome.json | 8 +- bin/submit_study.py | 1 - docs/CONTRIBUTING.md | 14 ++ {assets => docs/images}/seqsubmit_schema.svg | 0 docs/methods.md | 66 ++++---- docs/usage.md | 154 +----------------- main.nf | 6 +- nextflow.config | 1 - nextflow_schema.json | 11 +- ro-crate-metadata.json | 2 +- subworkflows/local/genome_evaluation/main.nf | 28 ++-- .../utils_nfcore_seqsubmit_pipeline/main.nf | 8 + workflows/assemblysubmit.nf | 29 ++-- workflows/genomesubmit.nf | 45 ++--- workflows/readsubmit.nf | 17 +- 17 files changed, 115 insertions(+), 288 deletions(-) rename {assets => docs/images}/seqsubmit_schema.svg (100%) diff --git a/README.md b/README.md index 41a1e7e..e3e201a 100644 --- a/README.md +++ b/README.md @@ -31,14 +31,13 @@ Currently, the pipeline supports four submission modes, each routed to a dedicat ![seqsubmit workflow diagram](docs/images/seqsubmit_schema.png) - **nf-core/seqsubmit** is a Nextflow pipeline for submitting sequence data to [ENA](https://www.ebi.ac.uk/ena/browser/home). The pipeline currently supports the following submission modes, each routed to a dedicated workflow: -- `reads` — raw sequencing reads submission via the `READSUBMIT` workflow (pink) -- `metagenomic_assemblies` — assembly submission via the `ASSEMBLYSUBMIT` workflow (green) -- `mags` — metagenome-assembled genomes (MAGs) submission via the `GENOMESUBMIT` workflow (blue) -- `bins` — bins submission via the `GENOMESUBMIT` workflow (blue) +- `reads` — raw sequencing reads submission via the `READSUBMIT` workflow (pink) +- `metagenomic_assemblies` — assembly submission via the `ASSEMBLYSUBMIT` workflow (green) +- `mags` — metagenome-assembled genomes (MAGs) submission via the `GENOMESUBMIT` workflow (blue) +- `bins` — bins submission via the `GENOMESUBMIT` workflow (blue) diff --git a/assets/schema_input_assembly.json b/assets/schema_input_assembly.json index 2b5fcea..6123501 100644 --- a/assets/schema_input_assembly.json +++ b/assets/schema_input_assembly.json @@ -77,13 +77,13 @@ "type": "string", "pattern": "^\\S+$", "errorMessage": "Assembler must be provided and cannot contain spaces", - "description": "Name of the assembler software used to generate the assembly" + "description": "Name of the assembler software used to generate the assembly, e.g. MEGAHIT, SPAdes, metaSPAdes, Flye, etc" }, "assembler_version": { "anyOf": [{ "type": "string" }, { "type": "number" }], "pattern": "^\\S+$", "errorMessage": "Assembler version must be provided and cannot contain spaces", - "description": "Version of the assembler software used to generate the assembly" + "description": "Version of the assembler software used to generate the assembly, e.g 1.0.0" } }, "required": ["sample", "fasta", "run_accession", "assembler", "assembler_version"], diff --git a/assets/schema_input_genome.json b/assets/schema_input_genome.json index 65472dc..a603b2c 100644 --- a/assets/schema_input_genome.json +++ b/assets/schema_input_genome.json @@ -59,15 +59,15 @@ }, "assembly_software": { "type": "string", - "description": "Tool name and version that was used to assemble data" + "description": "Tool name and version that was used to assemble data, e.g. MEGAHIT_v1.0, SPAdes_v4.0.0, metaSPAdes_v3.15.0" }, "binning_software": { "type": "string", - "description": "Tool name and version that was used to bin data" + "description": "Tool name and version that was used to bin data, e.g. MetaBAT2, VAMB, MaxBin2, CONCOCT, SemiBin2, COMEBin, etc" }, "binning_parameters": { "type": "string", - "description": "Arguments used to bin data different from default" + "description": "Arguments used to bin data different from default, e.g. -min_contig_length 1000" }, "stats_generation_software": { "type": "string", @@ -117,7 +117,7 @@ }, "NCBI_lineage": { "type": "string", - "description": "full NCBI lineage - format: x;y;z. For example, the lineage for E. coli can be: 'Bacteria;Pseudomonadati;Pseudomonadota;Gammaproteobacteria;Enterobacterales;Enterobacteriaceae;Escherichia' or '2;1224;1236;91347;543;561;562'" + "description": "full NCBI lineage - format: x;y;z. For example, the lineage for E. coli can be: 'Bacteria;Pseudomonadati;Pseudomonadota;Gammaproteobacteria;Enterobacterales;Enterobacteriaceae;Escherichia' or '2;1224;1236;91347;543;561;562'. For more info check https://www.ncbi.nlm.nih.gov/datasets/docs/v2/data-processing/taxonomy-processing/taxonomy/" } }, "required": [ diff --git a/bin/submit_study.py b/bin/submit_study.py index 23f43fd..6c8b0d4 100755 --- a/bin/submit_study.py +++ b/bin/submit_study.py @@ -379,7 +379,6 @@ def _add_project_element( sp = ET.SubElement(project, "SUBMISSION_PROJECT") ET.SubElement(sp, "SEQUENCING_PROJECT") - # TODO: Check existing_study_type and new_study_type metadata fields, do we need those? study_type = study.get("existing_study_type") if study_type: attrs = ET.SubElement( diff --git a/docs/CONTRIBUTING.md b/docs/CONTRIBUTING.md index 7552e65..2ff2980 100644 --- a/docs/CONTRIBUTING.md +++ b/docs/CONTRIBUTING.md @@ -184,6 +184,20 @@ If you update images or graphics, follow the nf-core [style guidelines](https:// To ensure consistency and maintainability of the nf-core/seqsubmit pipeline, contributors should follow these guidelines when implementing new features or modifying existing functionality. +### Code style + +The pipeline is organized into three separate workflows `(mode`s-) because each mode has distinct procedures and requirements. Future development should follow the same structure to maintain consistency and readability. + +If a new feature extends support for an **existing** data type (for example, _adding support for eukaryotic MAGs_), the corresponding logic should be integrated into the existing workflow rather than creating a new one. + +To improve readability, divide the main workflow into logical sections using comments that begin with `// ---------`, for example: + +``` +// --------- Upload data to ENA +``` + +Use standard `//` comments for shorter explanations within each section. + ### ENA Submission Requirements Submissions to [ENA](https://www.ebi.ac.uk/ena/browser/home) are governed by a well-defined data model and submission process. These include relationships between entities (e.g. studies, samples, experiments, runs, analyses), required submission steps, and accepted file types and formats. diff --git a/assets/seqsubmit_schema.svg b/docs/images/seqsubmit_schema.svg similarity index 100% rename from assets/seqsubmit_schema.svg rename to docs/images/seqsubmit_schema.svg diff --git a/docs/methods.md b/docs/methods.md index acbf12f..67e5488 100644 --- a/docs/methods.md +++ b/docs/methods.md @@ -2,6 +2,38 @@ This page provides a detailed overview of the methods and procedures that are executed throughout the nf-core/seqsubmit pipeline. +## `ASSEMBLYSUBMIT` + +### Overview + +The `ASSEMBLYSUBMIT` workflow: + +1. Reads the samplesheet and associated assembly FASTA files. +2. Validates assembly FASTA files. +3. Reuses provided or calculates missing `coverage` values. +4. Builds the metadata table used for manifest generation. +5. Generates assembly manifests and submits the assemblies to ENA. + +### Assembly FASTA validation + +Assembly FASTA files are validated with `fa-lint` before downstream processing. Each file is checked for FASTA format validity and contig count. A genome must contain at least two contigs to pass validation, which is an ENA requirement for contig-level submissions. + +Only assemblies with successful validation are forwarded to coverage estimation, metadata/manifest generation and submission. + +### Coverage calculation + +If the `coverage` column is already populated in the samplesheet, that value is used directly. + +If `coverage` is missing, the workflow calculates coverage with `coverm contig`. + +`coverm contig` outputs per-contig depth. The workflow then reads this file and calculates the arithmetic mean across all contigs. + +If the per-contig coverage values are $c_1, c_2, \ldots, c_n$, the workflow currently computes assembly coverage as an unweighted mean across contigs: + +$$ +\bar{c} = \frac{1}{n} \sum_{i=1}^{n} c_i +$$ + ## `GENOMESUBMIT` ### Overview @@ -93,7 +125,7 @@ If `NCBI_lineage` is already present in the input samplesheet, the value is reta Before classification, input FASTA files are normalized to a `.fasta` suffix by `RENAME_FASTA_FOR_CATPACK`. Classification is then run in bin mode (`CAT_pack bins`, followed by `CAT_pack add_names`). -Database input is taken from `--cat_db` when provided; otherwise the workflow uses `--cat_db_download_id` to download and prepare a CATPACK database. The resulting classification table is parsed, and the lineage field is written to `NCBI_lineage`. +Database input is taken from `--cat_db` when provided; otherwise the workflow uses `nr` database identifier to download and prepare a CATPACK NCBI database. See [CAT_pack documentation](https://github.com/MGXlab/CAT_pack#downloading-preconstructed-database-files). There are two options available: `nr` - NCBI non-redundant protein database and `gtdb` (Genome Taxonomy Database (GTDB) proteins). We use NCBI proteins because ENA supports only NCBI format of taxonomy for submission. The resulting classification table is parsed, and the lineage field is written to `NCBI_lineage`. ### Completeness and contamination assessment @@ -102,35 +134,3 @@ Completeness and contamination are evaluated together in a shared genome quality The workflow checks three samplesheet fields: `completeness`, `contamination`, and `stats_generation_software`. If all three are already present, those values are reused. If any of them is missing, the genome is analysed with `CheckM2 predict`. If `--checkm2_db` is supplied and exists, that database is used directly. Otherwise, the workflow downloads a CheckM2 database from Zenodo (using the configured database ID) and then runs prediction. For records that run `CheckM2`, completeness and contamination are extracted from the generated quality report (`quality_report.tsv`) and used `CheckM2` version is recorded as `stats_generation_software`. - -## `ASSEMBLYSUBMIT` - -### Overview - -The `ASSEMBLYSUBMIT` workflow: - -1. Reads the samplesheet and associated assembly FASTA files. -2. Validates assembly FASTA files. -3. Reuses provided or calculates missing `coverage` values. -4. Builds the metadata table used for manifest generation. -5. Generates assembly manifests and submits the assemblies to ENA. - -### Assembly FASTA validation - -Assembly FASTA files are validated with `fa-lint` before downstream processing. Each file is checked for FASTA format validity and contig count. A genome must contain at least two contigs to pass validation, which is an ENA requirement for contig-level submissions. - -Only assemblies with successful validation are forwarded to coverage estimation, metadata/manifest generation and submission. - -### Coverage calculation - -If the `coverage` column is already populated in the samplesheet, that value is used directly. - -If `coverage` is missing, the workflow calculates coverage with `coverm contig`. - -`coverm contig` outputs per-contig depth. The workflow then reads this file and calculates the arithmetic mean across all contigs. - -If the per-contig coverage values are $c_1, c_2, \ldots, c_n$, the workflow currently computes assembly coverage as an unweighted mean across contigs: - -$$ -\bar{c} = \frac{1}{n} \sum_{i=1}^{n} c_i -$$ diff --git a/docs/usage.md b/docs/usage.md index 17538e7..ea3a386 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -32,7 +32,7 @@ The pipeline supports the following data types, selected via the `--mode` parame - `mags` — metagenome-assembled genomes (MAGs) - `bins` — metagenomic bins -Each mode has its own samplesheet structure, prerequisites, and limitations — see the dedicated section for the mode you want to use below. +Each mode has its own samplesheet _structure_, _prerequisites_, and _limitations_ — see the dedicated section for the mode you want to use below. ## Before you start @@ -52,110 +52,7 @@ Before running the pipeline in any mode, make sure that: - Provide either a study accession or a study registration metadata file for the study the submission will be associated with. See the [Submission study](#submission-study) section below for details. -<<<<<<< HEAD -```bash ---input '[path to samplesheet.csv]' -``` - -### `mags` and `bins` modes (`GENOMESUBMIT`) - -Use this samplesheet structure for MAG and bin submission. The input format follows [assets/schema_input_genome.json](../assets/schema_input_genome.json). - -Example: - -```csv title="samplesheet_genomes.csv" -sample,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage -mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default,CheckM2 1.0.1,92.81,1.09,66.04,sediment metagenome,No,marine,cable bacteria,marine sediment,No,d__Bacteria;p__Proteobacteria;s__ -``` - -> [!IMPORTANT] -> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. - -| Column | Description | -| --------------------------- | ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | A unique identifier for this data entry. Must be globally unique within the input dataset. | -| `fasta` | Path to MAG/bin contigs in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. | -| `accession` | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | -| `fastq_1` | Path to the read file in FASTQ format used to generate the source metagenomic assembly. Required if `genome_coverage` is not provided. | -| `fastq_2` | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | -| `assembly_software` | Tool name and version that were used to generate the source metagenomic assembly. | -| `binning_software` | Binning tool, including version, that was used to generate the bins. | -| `binning_parameters` | Arguments that were used during binning. | -| `stats_generation_software` | Tool, including version, that was used to calculate completeness and contamination. | -| `completeness` | Genome completeness value. | -| `contamination` | Genome contamination value. | -| `genome_coverage` | Estimated average sequencing depth across the genome. If the value is missing, it is computed automatically during pipeline execution when reads are provided. | -| `metagenome` | Registered metagenome taxonomic identifier or name that matches an existing ENA taxonomy entry. For more details see https://ena-docs.readthedocs.io/en/latest/faq/taxonomy.html | -| `co-assembly` | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: Yes or No. | -| `broad_environment` | Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428) | -| `local_environment` | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". | -| `environmental_medium` | Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). | -| `RNA_presence` | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: Yes or No. | -| `NCBI_lineage` | NCBI taxonomy lineage of the genome. Can be composted of either numerical IDs or taxon names separated by ";". | - -> [!NOTE] -> More information about envioronment tags can be found at checklists [ERC000050](https://www.ebi.ac.uk/ena/browser/view/ERC000050) for bins and [ERC000047](https://www.ebi.ac.uk/ena/browser/view/ERC000047) for MAGs under the field names "broad-scale environmental context", "local environmental context", and "environmental medium". - -### `metagenomic_assemblies` mode (`ASSEMBLYSUBMIT`) - -Use this samplesheet structure for metagenomic assembly submission. The input format follows [assets/schema_input_assembly.json](../assets/schema_input_assembly.json). - -Provide either read files (`fastq_1`, optionally `fastq_2`) or a `coverage` value for each row. If `coverage` is missing and reads are provided, the workflow calculates average coverage automatically. - -Example: - -```csv title="samplesheet_assembly.csv" -sample,fasta,fastq_1,fastq_2,coverage,run_accession,assembler,assembler_version -assembly_001,data/assembly_001.fasta.gz,data/assembly_001_R1.fastq.gz,data/assembly_001_R2.fastq.gz,,ERR011322,SPAdes,3.15.5 -assembly_002,data/assembly_002.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 -``` - -> [!IMPORTANT] -> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. - -| Column | Description | -| ------------------- | ----------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | A unique identifier for this data entry. Must be globally unique within the input dataset. | -| `fasta` | Path to assembly contigs in FASTA format compressed with `gzip`. | -| `fastq_1` | Path to the read file in FASTQ format used to generate the metagenomic assembly. Required if `coverage` is not provided. | -| `fastq_2` | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | -| `coverage` | Estimated sequencing depth of the assembly. If this value is missing, it is computed automatically during pipeline execution when reads are provided. | -| `run_accession` | ENA run accession for the reads used to generate the metagenomic assembly. Reads must already be submitted to ENA. | -| `assembler` | Name of the assembler software used to generate the assembly. | -| `assembler_version` | Version of the assembler software used to generate the assembly. | - -An example file is available at [assets/samplesheet_assembly.csv](../assets/samplesheet_assembly.csv). - -### `reads` mode (`READSUBMIT`) - -Use this samplesheet structure for raw sequencing reads submission. The input format follows [assets/schema_input_reads.json](../assets/schema_input_reads.json). - -Example: - -```csv title="samplesheet_reads.csv" -sample,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,library_selection,library_strategy,insert_size,library_name,description -illumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLUMINA,Illumina HiSeq 2000,GENOMIC,RANDOM,WGS,500,HiSeq_library_001,Illumina sequencing of sample XYZ -pacbio_run_001,SAMEA7654321,data/pacbio_reads.fastq.gz,,PACBIO_SMRT,PacBio Sequel,GENOMIC,RANDOM,WGS,,PacBio_library_002,Long-read sequencing -``` - -> [!IMPORTANT] -> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. - -| Column | Type | Required | Description | -| ------------------- | --------- | -------- | --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | str | Yes | Unique identifier of this particular data entry. Used as an experiment name. | -| `sample_accession` | str | Yes | ENA sample accession (starting with SAMEA) of the sample used to generate raw reads. | -| `fastq_1` | file path | Yes | Path to forward reads in FASTQ format (optionally gzipped). | -| `fastq_2` | file path | No | Path to reverse reads for paired-end data. Leave empty for single-end reads. | -| `platform` | str | Yes | Sequencing platform. Supported values: `ILLUMINA`, `PACBIO_SMRT`, `OXFORD_NANOPORE`, `ION_TORRENT`, `CAPILLARY`, `DNBSEQ`, `ELEMENT`, `GENAPSYS`, `GENEMIND`, `HELICOS`, `LS454`, `BGISEQ`, `ULTIMA`, `VELA_DIAGNOSTICS`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | -| `instrument` | str | Yes | Sequencer model, e.g. "Illumina HiSeq 2000", "PacBio Sequel", "MinION". | -| `library_source` | str | Yes | Library source type. Options: `GENOMIC`, `METAGENOMIC`, `TRANSCRIPTOMIC`, `METAGENOMIC SINGLE CELL`, `TRANSCRIPTOMIC SINGLE CELL`, `SYNTHETIC`, `VIRAL RNA`, `OTHER`. | -| `library_selection` | str | Yes | Library selection method. Options: `RANDOM`, `PCR`, `RANDOM PCR`, `RT-PCR`, `MF`, `cDNA`, `cDNA_randomPriming`, `cDNA_oligo_dT`, `PolyA`, `Inverse rRNA`, `ChIP`, `MNase`, `DNase`, `Hybrid Selection`, etc. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | -| `library_strategy` | str | Yes | Library strategy. Options: `WGS`, `WGA`, `WXS`, `RNA-Seq`, `miRNA-Seq`, `ncRNA-Seq`, `EST`, `Hi-C`, `ATAC-seq`, `WCS`, `RAD-Seq`, `CLONE`, `AMPLICON`, `POOLCLONE`, `etc`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | -| `insert_size` | number | No | Fragment/insert size for paired-end reads (e.g., 500 for 500 bp inserts). Leave empty if not applicable. | -| `library_name` | str | No | Descriptive library name (optional). | -| `description` | str | No | Free-text description of the experiment (optional). | - +- Depending on the chosen mode, samples, reads, or metagenomic assemblies must be pre-submitted to ENA to obtain the corresponding accessions, which you then reference in your submission. Refer to the relevant mode section below for details. ## Submission study @@ -449,7 +346,7 @@ In particular, the CAT_pack database is extremely large and may take a long time - provide the path to a local database with `--checkm2_db`, otherwise the pipeline downloads the version specified with `--checkm2_db_download_id` (by default Zenodo accession `14897628`). - CAT_pack: - - provide the path to a local database (containing `tax/` and `db/` folders or a `tar.gz` archive) with `--cat_db`, otherwise the pipeline constructs the version specified with `--cat_db_download_id` (by default `nr`). + - provide the path to a local database (containing `tax/` and `db/` folders or a `tar.gz` archive) with `--cat_db`, otherwise the pipeline constructs NCBI non-redundant protein database. See the [CAT_pack documentation](https://github.com/MGXlab/CAT_pack) and [CheckM2 documentation](https://github.com/chklovski/CheckM2) for more details on usage and creation of databases. @@ -464,7 +361,7 @@ In addition to the [common parameters](#running-the-pipeline): | -------------- | -------- | ---------------------------------------------------------------------------------------------------------------------------------------------- | | `--upload_tpa` | No | Mark assemblies as [third party assemblies (TPA)](#third-party-assemblies-tpa) when required. Default: `false`. | | `--checkm2_db` | No | Path to a local CheckM2 database. If omitted, downloads the version set by `--checkm2_db_download_id`. Default: `14897628` (Zenodo accession). | -| `--cat_db` | No | Path to a local CAT_pack database. If omitted, constructs the version set by `--cat_db_download_id`. Default: `nr`. | +| `--cat_db` | No | Path to a local CAT_pack database. If omitted, constructs the NCBI non-redundant protein database. | Example `mags` mode upload to TEST server, run command with docker: @@ -530,50 +427,7 @@ Parameters common to all modes: | `--is_private` | No | Use if you are referencing private data accessions in your submission. Default: `false`. | | `--release_date` | No | Use if you want to keep your data private after submission until a particular date. | -<<<<<<< HEAD -Example `mags` mode upload to TEST server, run command with docker: - -```bash -nextflow run nf-core/seqsubmit \ - -profile docker \ - --mode mags \ - --input assets/samplesheet_genomes.csv \ - --submission_study \ - --centre_name TEST_CENTER \ - --webincli_mode submit \ - --test_upload \ - --outdir results/validate_mags -``` - -Example `metagenomic_assemblies` mode upload to TEST server, run command with docker: - -```bash -nextflow run nf-core/seqsubmit \ - -profile docker \ - --mode metagenomic_assemblies \ - --input assets/samplesheet_assembly.csv \ - --submission_study \ - --centre_name TEST_CENTER \ - --webincli_mode submit \ - --test_upload \ - --outdir results/validate_assemblies -``` - -Example `reads` mode upload to TEST server, run command with docker: - -```bash -nextflow run nf-core/seqsubmit \ - -profile docker \ - --mode reads \ - --input samplesheet_reads.csv \ - --submission_study \ - --webincli_mode submit \ - --test_upload \ - --outdir results/validate_reads -``` -======= Each mode also has additional parameters and example commands — see its own "`` parameters and example" subsection above. ->>>>>>> 1.0.0 If you wish to repeatedly use the same parameters for multiple runs, rather than specifying each flag in the command, you can specify these in a params file. diff --git a/main.nf b/main.nf index f741919..2ba497e 100644 --- a/main.nf +++ b/main.nf @@ -55,7 +55,7 @@ workflow NFCORE_SEQSUBMIT { params.checkm2_db, params.checkm2_db_download_id, params.cat_db, - params.cat_db_download_id, + "nr", // NCBI non-redundant protein database identifier params.centre_name, params.upload_tpa, params.test_upload, @@ -123,7 +123,9 @@ workflow { params.mode, params.help, params.help_full, - params.show_hidden + params.show_hidden, + params.submission_study, + params.study_metadata ) // diff --git a/nextflow.config b/nextflow.config index 2cef81c..c4a1fba 100644 --- a/nextflow.config +++ b/nextflow.config @@ -31,7 +31,6 @@ params { // NCBI taxonomy cat_db = null - cat_db_download_id = 'nr' // MultiQC options multiqc_config = null diff --git a/nextflow_schema.json b/nextflow_schema.json index a8bf46c..d6323b9 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -71,17 +71,10 @@ "help_text": "Provide a path to pre-downloaded DB or DB id for automatic download", "properties": { "cat_db": { - "type": "file-path", + "format": "file-path", "description": "Path to local CAT_pack DB", "help": "Database should be pre-downloaded using CATpack commands. The folder with database should contain sub-folders 'db' with files .dmnd, .fastaid2LCAtaxid, .taxids_with_multiple_offspring, etc, as well as subfolder 'tax' that should contain taxonomy files like .dmp, .taxdump.tar.gz, .prot.accession2taxid.FULL.gz, etc", "fa_icon": "fas fa-users-cog" - }, - "cat_db_download_id": { - "type": "string", - "description": "CAT_pack database ID for download", - "help": "There are two options available: nr (default) NCBI non-redundant protein database and gtdb (Genome Taxonomy Database (GTDB) proteins). We use NCBI proteins because ENA supports only NCBI format of taxonomy for submission", - "fa_icon": "fas fa-users-cog", - "default": "nr" } } }, @@ -93,7 +86,7 @@ "help_text": "These options are used to run Checkm2 that will calculate completeness and contamination of genome if those were not provided as input", "properties": { "checkm2_db": { - "type": "file-path", + "format": "file-path", "description": "Path to pre-downloaded checkm2 database", "fa_icon": "fas fa-users-cog" }, diff --git a/ro-crate-metadata.json b/ro-crate-metadata.json index d420ccc..7b73754 100644 --- a/ro-crate-metadata.json +++ b/ro-crate-metadata.json @@ -23,7 +23,7 @@ "@type": "Dataset", "creativeWorkStatus": "InProgress", "datePublished": "2026-04-30T13:32:49+00:00", - "description": "

    \n \n \n \"nf-core/seqsubmit\"\n \n

    \n\n[![Open in GitHub Codespaces](https://img.shields.io/badge/Open_In_GitHub_Codespaces-black?labelColor=grey&logo=github)](https://github.com/codespaces/new/nf-core/seqsubmit)\n[![GitHub Actions CI Status](https://github.com/nf-core/seqsubmit/actions/workflows/nf-test.yml/badge.svg)](https://github.com/nf-core/seqsubmit/actions/workflows/nf-test.yml)\n[![GitHub Actions Linting Status](https://github.com/nf-core/seqsubmit/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/seqsubmit/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/seqsubmit/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX)\n[![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com)\n\n[![Nextflow](https://img.shields.io/badge/version-%E2%89%A525.10.4-green?style=flat&logo=nextflow&logoColor=white&color=%230DC09D&link=https%3A%2F%2Fnextflow.io)](https://www.nextflow.io/)\n[![nf-core template version](https://img.shields.io/badge/nf--core_template-4.0.2-green?style=flat&logo=nfcore&logoColor=white&color=%2324B064&link=https%3A%2F%2Fnf-co.re)](https://github.com/nf-core/tools/releases/tag/4.0.2)\n[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/)\n[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/)\n[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/)\n[![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/seqsubmit)\n\n[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23seqsubmit-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/seqsubmit)[![Follow on Bluesky](https://img.shields.io/badge/bluesky-%40nf__core-1185fe?labelColor=000000&logo=bluesky)](https://bsky.app/profile/nf-co.re)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core)\n\n## Introduction\n\n**nf-core/seqsubmit** is a Nextflow pipeline for submitting sequence data to [ENA](https://www.ebi.ac.uk/ena/browser/home).\nCurrently, the pipeline supports four submission modes, each routed to a dedicated workflow and requiring its own input samplesheet structure:\n\n- `mags` for Metagenome Assembled Genomes (MAGs) submission with `GENOMESUBMIT` workflow\n- `bins` for bins submission with `GENOMESUBMIT` workflow\n- `metagenomic_assemblies` for assembly submission with `ASSEMBLYSUBMIT` workflow\n- `reads` for raw sequencing reads submission with `READSUBMIT` workflow\n\n![seqsubmit workflow diagram](assets/seqsubmit_schema.png)\n\n## Requirements\n\n- [Nextflow](https://www.nextflow.io/) `>=25.04.0`\n- Webin account registered at https://www.ebi.ac.uk/ena/submit/webin/login\n- Raw reads used to assemble contigs submitted to [INSDC](https://www.insdc.org/) and associated accessions available\n\nSetup your environment secrets before running the pipeline:\n\n`nextflow secrets set ENA_WEBIN \"Webin-XXX\"`\n\n`nextflow secrets set ENA_WEBIN_PASSWORD \"XXX\"`\n\nMake sure you update commands above with your authorised credentials.\n\n## Input samplesheets\n\nFor detailed descriptions of all samplesheet columns, see the [usage documentation](docs/usage.md#samplesheet-input).\n\n### `mags` and `bins` modes (`GENOMESUBMIT`)\n\nThe input must follow `assets/schema_input_genome.json`.\n\nRequired columns:\n\n- `sample`\n- `fasta` (must end with `.fa.gz`, `.fasta.gz`, or `.fna.gz`)\n- `accession`\n- `assembly_software`\n- `binning_software`\n- `binning_parameters`\n- `metagenome`\n- `environmental_medium`\n- `broad_environment`\n- `local_environment`\n- `co-assembly`\n\nAt least one of the following must be provided per row:\n\n- reads (`fastq_1`, optional `fastq_2` for paired-end)\n- `genome_coverage`\n\nAdditional supported columns:\n\n- `stats_generation_software`\n- `completeness`\n- `contamination`\n- `RNA_presence`\n- `NCBI_lineage`\n\nIf `genome_coverage`, `stats_generation_software`, `completeness`, `contamination`, `RNA_presence`, or `NCBI_lineage` are missing, the workflow can calculate or infer them when the required inputs are available.\n\nThose fields are metadata required for the [genome_uploader](https://github.com/EBI-Metagenomics/genome_uploader) package.\n\nExample `samplesheet_genomes.csv`:\n\n```csv\nsample,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage\nlachnospira_eligens,data/bin_lachnospira_eligens.fa.gz,SRR24458089,,,spades_v3.15.5,metabat2_v2.6,default,CheckM2_v1.0.1,61.0,0.21,32.07,sediment metagenome,No,marine,cable_bacteria,marine_sediment,No,d__Bacteria;p__Proteobacteria;s__unclassified_Proteobacteria\n```\n\n> [!IMPORTANT]\n> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown.\n\n### `metagenomic_assemblies` mode (`ASSEMBLYSUBMIT`)\n\nThe input must follow `assets/schema_input_assembly.json`.\n\nRequired columns:\n\n- `sample`\n- `fasta` (must end with `.fa.gz`, `.fasta.gz`, or `.fna.gz`)\n- `run_accession`\n- `assembler`\n- `assembler_version`\n\nAt least one of the following must be provided per row:\n\n- reads (`fastq_1`, optional `fastq_2` for paired-end)\n- `coverage`\n\nIf `coverage` is missing and reads are provided, the workflow calculates average coverage with `coverm`.\n\nExample `samplesheet_assembly.csv`:\n\n```csv\nsample,fasta,fastq_1,fastq_2,coverage,run_accession,assembler,assembler_version\nassembly_1,data/contigs_1.fasta.gz,data/reads_1.fastq.gz,data/reads_2.fastq.gz,,ERR011322,SPAdes,3.15.5\nassembly_2,data/contigs_2.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9\n```\n\n> [!IMPORTANT]\n> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown.\n\n### `reads` mode (`READSUBMIT`)\n\nThe input must follow `assets/schema_input_reads.json`.\n\nRequired columns:\n\n- `sample`\n- `sample_accession`\n- `fastq_1`\n- `fastq_2`\n- `platform`\n- `instrument`\n- `library_source`\n- `library_selection`\n- `library_strategy`\n\nOptional columns:\n\n- `insert_size`\n- `library_name`\n- `description`\n\nExample `samplesheet_reads.csv`:\n\n```csv\nsample,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,library_selection,library_strategy,insert_size,library_name,description\nillumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLUMINA,Illumina HiSeq 2000,GENOMIC,RANDOM,WGS,500,HiSeq_library_001,Illumina sequencing of sample XYZ\n```\n\n> [!IMPORTANT]\n> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown.\n\n## Usage\n\n> [!NOTE]\n> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/get_started/environment_setup/overview) on how to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/get_started/run-your-first-pipeline) with `-profile test` before running the workflow on actual data.\n\n### Submission study\n\nAll data submitted through this pipeline must be associated with an ENA study (project). You can either pass an accession of your existing study via `--submission_study`or provide a metadata file via `--study_metadata` and the pipeline will register the study with ENA before submitting your data.\n\nSee the [usage documentation](docs/usage.md#submission-study) for more details.\n\n### Data privacy\n\nYou can reference private ENA data if you have access to it via your Webin account (it was submitted previously under your credentials). To use private data accessions in your submission metadata, specify the `--is_private` flag. The pipeline will then use your provided Webin credentials to fetch the required metadata.\n\nYou can also control the privacy of your submitted data:\n\n- If you provide an existing study via `--submission_study`, your submission will inherit the same privacy status as that study.\n- If no study is provided, the pipeline will register a new one for you. To keep this new study private, you must specify a release date using `--release_date YYYY-MM-DD` (up to 2 years from the current date).\n\n| Example | Source Data | Submission Visibility | Required Arguments | Result |\n| --------------------------------------------------------------------------------------------------------------------------------------------------------------------- | ----------- | --------------------- | ------------------------------------------------------ | ----------------------------------------------------- |\n| Assemblies generated from public reads. Assemblies should be public immediately after submission | public | public | \u2013 | SUCCESS |\n| Assemblies generated from public reads. Assemblies should remain private for 1 year after submission | public | private | `--release_date YYYY-MM-DD` (+1 year) | SUCCESS |\n| Bins generated from private reads and assemblies. Bins should be public after submission. User **has access** to the reads and assemblies via Webin account | private | public | `--is_private` | SUCCESS |\n| Bins generated from private reads and assemblies. Bins should remain private for 2 years after submission. User **has access** via Webin account | private | private | `--is_private`, `--release_date YYYY-MM-DD` (+2 years) | SUCCESS |\n| MAGs generated from private reads and assemblies. MAGs should be public after submission. User **does not have access** to the reads and assemblies via Webin account | private | public | \u2013 | ERROR (submission can only reference accessible data) |\n\n### Database setup (`CheckM2` and `CAT_pack`)\n\nThe `mags`/`bins` workflow requires databases for completeness/contamination estimation and taxonomy assignment. See [Usage documentation](usage.md) for details.\n\n### Required parameters:\n\n| Parameter | Description |\n| ------------------------------------------ | ----------------------------------------------------------------------------------------------------------------- |\n| `--mode` | Type of the data to be submitted. Options: `[mags, bins, metagenomic_assemblies, reads]` |\n| `--input` | Path to the samplesheet describing the data to be submitted |\n| `--outdir` | Path to the output directory for pipeline results |\n| `--submission_study` OR `--study_metadata` | ENA study accession (PRJ/ERP) to submit the data to OR metadata file in JSON/TSV/CSV format to register new study |\n| `--centre_name` | Name of the submitter's organisation |\n\n### Optional parameters:\n\n| Parameter | Description |\n| ----------------- | ------------------------------------------------------------------------------------------ |\n| `--upload_tpa` | Flag to control the type of assembly study (third party assembly or not). Default: false |\n| `--test_upload` | Upload to TEST ENA server instead of LIVE. Default: true |\n| `--webincli_mode` | Choose Webin-CLI mode: `submit` or `validate`. Default: `submit` |\n| `--private` | Use that flag if you are referring to private data in ENA |\n| `--release_date` | Use that flag if you want to keep your data private after submission until particular date |\n\nGeneral command template:\n\n```bash\nnextflow run nf-core/seqsubmit \\\n -profile \\\n --mode \\\n --input \\\n --centre_name \\\n --submission_study \\\n --outdir \n```\n\nTest run (submission to the ENA TEST server) in `mags` mode:\n\n```bash\nnextflow run nf-core/seqsubmit \\\n -profile docker \\\n --mode mags \\\n --input assets/samplesheet_genomes.csv \\\n --submission_study \\\n --centre_name TEST_CENTER \\\n --webincli_mode submit \\\n --test_upload true \\\n --outdir results/validate_mags\n```\n\nTest run (submission to the ENA TEST server) in `metagenomic_assemblies` mode:\n\n```bash\nnextflow run nf-core/seqsubmit \\\n -profile docker \\\n --mode metagenomic_assemblies \\\n --input assets/samplesheet_assembly.csv \\\n --submission_study \\\n --centre_name TEST_CENTER \\\n --webincli_mode submit \\\n --test_upload true \\\n --outdir results/validate_assemblies\n```\n\nLive submission example:\n\n```bash\nnextflow run nf-core/seqsubmit \\\n -profile docker \\\n --mode metagenomic_assemblies \\\n --input assets/samplesheet_assembly.csv \\\n --submission_study PRJEB98843 \\\n --test_upload false \\\n --webincli_mode submit \\\n --outdir results/live_assembly\n```\n\n> [!WARNING]\n> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; see [docs](https://nf-co.re/docs/running/run-pipelines#using-parameter-files).\n\nFor more details and further functionality, please refer to the [usage documentation](https://nf-co.re/seqsubmit/usage) and the [parameter documentation](https://nf-co.re/seqsubmit/parameters).\n\n## Pipeline output\n\nKey output locations in `--outdir`:\n\n- `mags/` or `bins/`: genome metadata, manifests, and per-sample submission support files\n- `metagenomic_assemblies/`: assembly metadata CSVs and per-sample coverage files\n- `multiqc/`: MultiQC summary report\n- `pipeline_info/`: execution reports, trace, DAG, and software versions\n\nFor full details, see the [output documentation](https://nf-co.re/seqsubmit/output).\n\n## Credits\n\nnf-core/seqsubmit was originally written by [Martin Beracochea](https://github.com/mberacochea), [Ekaterina Sakharova](https://github.com/KateSakharova), [Sofia Ochkalova](https://github.com/ochkalova), [Evangelos Karatzas](https://github.com/vagkaratzas) and [Tim Rozday](https://github.com/timrozday-mgnify).\n\n## Contributions and Support\n\nIf you would like to contribute to this pipeline, please see the [contributing guidelines](docs/CONTRIBUTING.md).\n\nFor further information or help, don't hesitate to get in touch on the [Slack `#seqsubmit` channel](https://nfcore.slack.com/channels/seqsubmit) (you can join with [this invite](https://nf-co.re/join/slack)).\n\n## Citations\n\n\n\n\n\nIf you use this pipeline please make sure to cite all used software.\nThis pipeline uses code and infrastructure developed and maintained by the [nf-core](https://nf-co.re) community, reused here under the [MIT license](https://github.com/nf-core/tools/blob/main/LICENSE).\n\n> **MGnify: the microbiome sequence data analysis resource in 2023**\n>\n> Richardson L, Allen B, Baldi G, Beracochea M, Bileschi ML, Burdett T, et al.\n>\n> Vol. 51, Nucleic Acids Research. Oxford University Press (OUP); 2022. p. D753\u20139. Available from: http://dx.doi.org/10.1093/nar/gkac1080\n\nAn extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file.\n\nYou can cite the `nf-core` publication as follows:\n\n> **The nf-core framework for community-curated bioinformatics pipelines.**\n>\n> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen.\n>\n> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x).\n", + "description": "

    \n \n \n \"nf-core/seqsubmit\"\n \n

    \n\n[![Open in GitHub Codespaces](https://img.shields.io/badge/Open_In_GitHub_Codespaces-black?labelColor=grey&logo=github)](https://github.com/codespaces/new/nf-core/seqsubmit)\n[![GitHub Actions CI Status](https://github.com/nf-core/seqsubmit/actions/workflows/nf-test.yml/badge.svg)](https://github.com/nf-core/seqsubmit/actions/workflows/nf-test.yml)\n[![GitHub Actions Linting Status](https://github.com/nf-core/seqsubmit/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/seqsubmit/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/seqsubmit/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX)\n[![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com)\n\n[![Nextflow](https://img.shields.io/badge/version-%E2%89%A525.10.4-green?style=flat&logo=nextflow&logoColor=white&color=%230DC09D&link=https%3A%2F%2Fnextflow.io)](https://www.nextflow.io/)\n[![nf-core template version](https://img.shields.io/badge/nf--core_template-4.0.2-green?style=flat&logo=nfcore&logoColor=white&color=%2324B064&link=https%3A%2F%2Fnf-co.re)](https://github.com/nf-core/tools/releases/tag/4.0.2)\n[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/)\n[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/)\n[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/)\n[![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/seqsubmit)\n\n[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23seqsubmit-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/seqsubmit)[![Follow on Bluesky](https://img.shields.io/badge/bluesky-%40nf__core-1185fe?labelColor=000000&logo=bluesky)](https://bsky.app/profile/nf-co.re)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core)\n\n## Introduction\n\n**nf-core/seqsubmit** is a Nextflow pipeline for submitting sequence data to [ENA](https://www.ebi.ac.uk/ena/browser/home).\nCurrently, the pipeline supports four submission modes, each routed to a dedicated workflow and requiring its own input samplesheet structure:\n\n- `mags` for Metagenome Assembled Genomes (MAGs) submission with `GENOMESUBMIT` workflow\n- `bins` for bins submission with `GENOMESUBMIT` workflow\n- `metagenomic_assemblies` for assembly submission with `ASSEMBLYSUBMIT` workflow\n- `reads` for raw sequencing reads submission with `READSUBMIT` workflow\n\n![seqsubmit workflow diagram](docs/images/seqsubmit_schema.png)\n\n**nf-core/seqsubmit** is a Nextflow pipeline for submitting sequence data to [ENA](https://www.ebi.ac.uk/ena/browser/home).\nThe pipeline currently supports the following submission modes, each routed to a dedicated workflow:\n\n- `reads` \u2014 raw sequencing reads submission via the `READSUBMIT` workflow (pink)\n- `metagenomic_assemblies` \u2014 assembly submission via the `ASSEMBLYSUBMIT` workflow (green)\n- `mags` \u2014 metagenome-assembled genomes (MAGs) submission via the `GENOMESUBMIT` workflow (blue)\n- `bins` \u2014 bins submission via the `GENOMESUBMIT` workflow (blue)\n\n\n\nEach workflow has its own samplesheet structure, prerequisites, and limitations \u2014 they are briefly described below. See the [usage documentation](https://nf-co.re/seqsubmit/usage) for more detailed explanations.\n\n## Requirements\n\n- [Nextflow](https://www.nextflow.io/) `>=25.04.0`\n- A Webin account registered at \n\n Set your Webin credentials as Nextflow secrets:\n\n ```bash\n nextflow secrets set ENA_WEBIN \"Webin-XXX\"\n nextflow secrets set ENA_WEBIN_PASSWORD \"XXX\"\n ```\n\n Make sure to replace the values above with your own credentials.\n\n- Provide either a study accession or a study registration metadata file for the study the submission will be associated with. See the [Submission study](docs/usage.md#submission-study) section of the usage documentation for details.\n\n- Depending on the chosen mode, samples, reads, or metagenomic assemblies must be pre-submitted to ENA to obtain the corresponding accessions, which you then reference in your submission. Refer to the relevant mode section in the [usage documentation](https://nf-co.re/seqsubmit/usage) for details.\n\n## Input samplesheets\n\n### `reads` mode\n\nExample:\n\n```csv\nsample,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,library_selection,library_strategy,insert_size,library_name,description\nillumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLUMINA,Illumina HiSeq 2000,GENOMIC,RANDOM,WGS,500,HiSeq_library_001,Illumina sequencing of sample XYZ\n```\n\nSee the [`reads` mode section](docs/usage.md#samplesheet-input) of the usage documentation for more details.\n\n### `metagenomic_assemblies` mode\n\nExample:\n\n```csv\nsample,fasta,fastq_1,fastq_2,coverage,run_accession,assembler,assembler_version\nassembly_1,data/contigs_1.fasta.gz,data/reads_1.fastq.gz,data/reads_2.fastq.gz,,ERR011322,SPAdes,3.15.5\nassembly_2,data/contigs_2.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9\n```\n\nSee the [`metagenomic_assemblies` mode section](docs/usage.md#samplesheet-input-1) of the usage documentation for more details.\n\n### `mags` and `bins` modes\n\nExample:\n\n```csv\nsample,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage\nlachnospira_eligens,data/bin_lachnospira_eligens.fa.gz,SRR24458089,,,spades_v3.15.5,metabat2_v2.6,default,CheckM2_v1.0.1,61.0,0.21,32.07,sediment metagenome,No,marine,cable_bacteria,marine_sediment,No,d__Bacteria;p__Proteobacteria;s__unclassified_Proteobacteria\n```\n\nSee the [`mags` and `bins` modes section](docs/usage.md#samplesheet-input-2) of the usage documentation for the full list of required and optional columns.\n\n## Usage\n\n> [!NOTE]\n> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/get_started/environment_setup/overview) on how to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/get_started/run-your-first-pipeline) with `-profile test` before running the workflow on actual data.\n\n### Running the pipeline\n\nEach mode also has its own additional parameters and example commands \u2014 see the [usage documentation](https://nf-co.re/seqsubmit/usage) for details. General command template:\n\n```bash\nnextflow run nf-core/seqsubmit \\\n -profile \\\n --mode \\\n --input \\\n --centre_name \\\n --submission_study \\\n --outdir \n```\n\n> [!WARNING]\n> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; see [docs](https://nf-co.re/docs/running/run-pipelines#using-parameter-files).\n\nFor more details and further functionality, please refer to the [usage documentation](https://nf-co.re/seqsubmit/usage) and the [parameter documentation](https://nf-co.re/seqsubmit/parameters).\n\n## Pipeline output\n\nKey output locations in `--outdir`:\n\n- `reads/`: per-sample submission receipts and accessions\n- `metagenomic_assemblies/`: assembly metadata CSVs and per-sample coverage files\n- `mags/` or `bins/`: genome metadata, manifests, and per-sample submission support files\n- `multiqc/`: MultiQC summary report\n- `pipeline_info/`: execution reports, trace, DAG, and software versions\n\nFor full details, see the [output documentation](https://nf-co.re/seqsubmit/output).\n\n## Credits\n\nnf-core/seqsubmit was originally written by [Martin Beracochea](https://github.com/mberacochea), [Ekaterina Sakharova](https://github.com/KateSakharova), [Sofia Ochkalova](https://github.com/ochkalova), [Evangelos Karatzas](https://github.com/vagkaratzas) and [Tim Rozday](https://github.com/timrozday-mgnify).\n\n## Contributions and Support\n\nIf you would like to contribute to this pipeline, please see the [contributing guidelines](docs/CONTRIBUTING.md).\n\nFor further information or help, don't hesitate to get in touch on the [Slack `#seqsubmit` channel](https://nfcore.slack.com/channels/seqsubmit) (you can join with [this invite](https://nf-co.re/join/slack)).\n\n## Citations\n\n\n\n\n\nIf you use this pipeline please make sure to cite all used software.\nThis pipeline uses code and infrastructure developed and maintained by the [nf-core](https://nf-co.re) community, reused here under the [MIT license](https://github.com/nf-core/tools/blob/main/LICENSE).\n\n> **MGnify: the microbiome sequence data analysis resource in 2023**\n>\n> Richardson L, Allen B, Baldi G, Beracochea M, Bileschi ML, Burdett T, et al.\n>\n> Vol. 51, Nucleic Acids Research. Oxford University Press (OUP); 2022. p. D753\u20139. Available from: http://dx.doi.org/10.1093/nar/gkac1080\n\nAn extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file.\n\nYou can cite the `nf-core` publication as follows:\n\n> **The nf-core framework for community-curated bioinformatics pipelines.**\n>\n> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen.\n>\n> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x).\n", "hasPart": [ { "@id": "main.nf" diff --git a/subworkflows/local/genome_evaluation/main.nf b/subworkflows/local/genome_evaluation/main.nf index 4f4921f..4c477ee 100644 --- a/subworkflows/local/genome_evaluation/main.nf +++ b/subworkflows/local/genome_evaluation/main.nf @@ -20,11 +20,9 @@ include { CHECKM2_PREDICT } from '../../../modules/nf-core/checkm2/pred workflow GENOME_EVALUATION { take: - ch_fasta // channel: [ val(meta), path(fasta) ] - ch_checkm2_db // channel: [ val(meta), path(db) ] - pre-built db as directory - // provide channel.empty() to trigger automatic download via ch_checkm2_db_download_id - ch_checkm2_db_download_id // channel: [ val(meta), val(db_id) ] - db ID for CHECKM2_DATABASEDOWNLOAD (e.g. '1234567') - // only used if ch_checkm2_db is empty + ch_fasta // channel: [ val(meta), path(fasta) ] + ch_checkm2_db // val: path to CheckM2 database or null + ch_checkm2_db_download_id // val: CheckM2 database download ID main: @@ -33,18 +31,12 @@ workflow GENOME_EVALUATION { // // Download and prepare db from scratch if no pre-built db provided - // Only trigger if ch_fasta has items - ch_download_trigger = ch_checkm2_db - .count() - .filter { count -> count == 0 } // Only proceed if ch_checkm2_db is empty - .combine(ch_fasta.first()) - .combine(ch_checkm2_db_download_id) - .map { _count, _meta, _fasta, db_meta, db_id -> [db_meta, db_id] } - - CHECKM2_DATABASEDOWNLOAD(ch_download_trigger) - - // Combine db sources - one of these channels will be empty depending on inputs - ch_db = ch_checkm2_db.mix(CHECKM2_DATABASEDOWNLOAD.out.database).first() + if (ch_checkm2_db) { + ch_checkm2_db = channel.value([[id: 'checkm2_db'], file(params.checkm2_db, checkIfExists: true)]) + } else { + CHECKM2_DATABASEDOWNLOAD(ch_checkm2_db_download_id) + ch_checkm2_db = CHECKM2_DATABASEDOWNLOAD.out.database + } // // Genome evaluation @@ -52,7 +44,7 @@ workflow GENOME_EVALUATION { CHECKM2_PREDICT( ch_fasta, - ch_db, + ch_checkm2_db, ) emit: diff --git a/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf b/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf index f65014a..4ea8248 100644 --- a/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf @@ -35,6 +35,8 @@ workflow PIPELINE_INITIALISATION { help // boolean: Display help message and exit help_full // boolean: Show the full help message show_hidden // boolean: Show hidden parameters in the help message + submission_study // val: ENA study accession for all reads in this run (optional). + study_metadata // val: path to study metadata file for study creation (used if no submission_study provided) main: @@ -113,6 +115,12 @@ workflow PIPELINE_INITIALISATION { ch_samplesheet = channel .fromList(samplesheetToList(input, "${projectDir}/assets/schema_input_reads.json")) } + + // check submission study and metadata presence/absence for REGISTER STUDY + if (!submission_study && !study_metadata) { + error("Either --submission_study or --study_metadata must be provided") + } + emit: samplesheet = ch_samplesheet versions = ch_versions diff --git a/workflows/assemblysubmit.nf b/workflows/assemblysubmit.nf index d599b59..b209609 100644 --- a/workflows/assemblysubmit.nf +++ b/workflows/assemblysubmit.nf @@ -48,7 +48,7 @@ workflow ASSEMBLYSUBMIT { def ch_versions = channel.empty() def ch_multiqc_files = channel.empty() - // Create assembly channel with proper metadata structure + // --------- Create assembly channel with proper metadata structure assembly_fasta = ch_samplesheet .map { row -> def meta = [ @@ -62,6 +62,7 @@ workflow ASSEMBLYSUBMIT { [meta, file(row[1])] } + // --------- Create reads channel with proper metadata structure reads_fastq = ch_samplesheet .filter { row -> row[2] && row[2] != "" } // Check if fastq_1 exists and is not empty .map { row -> @@ -88,15 +89,15 @@ workflow ASSEMBLYSUBMIT { assembly_fasta ) + // --------- Assembly coverage calculation // For assemblies without coverage, calculate coverage with CoverM - FASTA_VALIDATION.out.valid_fastas + coverm_input = FASTA_VALIDATION.out.valid_fastas .filter { meta, _fasta -> meta.coverage == null } .join(reads_fastq) .multiMap { meta, fasta, fastq -> assembly: [ meta, fasta ] reads: [ meta, fastq ] } - .set { coverm_input } COVERM_CONTIG ( coverm_input.reads, @@ -119,7 +120,7 @@ workflow ASSEMBLYSUBMIT { } // Update metadata with calculated coverage - FASTA_VALIDATION.out.valid_fastas + assemblies_with_added_cov_ch = FASTA_VALIDATION.out.valid_fastas .filter { meta, _fasta -> meta.coverage == null } .join( average_coverage_ch ) .map { meta, fasta, avg_coverage -> @@ -127,7 +128,6 @@ workflow ASSEMBLYSUBMIT { updated_meta.coverage = avg_coverage [updated_meta, fasta] } - .set { assemblies_with_added_cov_ch } // Combine assemblies with updated metadata (for samples that had coverage calculated) // and assemblies that already had coverage @@ -135,20 +135,18 @@ workflow ASSEMBLYSUBMIT { .filter { meta, _fasta -> meta.coverage != null } .mix( assemblies_with_added_cov_ch ) - // Create CSV with assembly metadata for manifest generation + // --------- Create CSV with assembly metadata for manifest generation CREATE_ASSEMBLY_METADATA_CSV( assemblies_with_coverage ) - // Concatenate assembly metadata CSVs into single file to publish + // --------- Concatenate assembly metadata CSVs into single file to publish CONCAT_METADATA ( CREATE_ASSEMBLY_METADATA_CSV.out.csv.map { _meta, file -> file }.collect().map { files -> [ [id: "assemblies_metadata"], files ] }, 'true' // skip_header - we want to keep the header from the first file and skip it for the rest ) - if (!submission_study && !study_metadata) { - error("Either --submission_study or --study_metadata must be provided") - } + // --------- Register study if accession was not provided via --submission_study def study_accession_ch if (submission_study) { // Use provided study accession directly @@ -168,7 +166,7 @@ workflow ASSEMBLYSUBMIT { } } - // Generate assembly manifest files and submit them to ENA + // --------- Generate assembly manifest files and submit them to ENA GENERATE_ASSEMBLY_MANIFEST( assemblies_with_coverage.join(CREATE_ASSEMBLY_METADATA_CSV.out.csv), study_accession_ch.first(), @@ -177,6 +175,7 @@ workflow ASSEMBLYSUBMIT { is_private ) + // --------- Upload data to ENA SUBMIT ( assemblies_with_coverage.join(GENERATE_ASSEMBLY_MANIFEST.out.manifest), test_upload, @@ -190,9 +189,7 @@ workflow ASSEMBLYSUBMIT { 'true' // skip_header - we want to keep the header from the first file and skip it for the rest ) - // - // Collate and save software versions - // + // --------- Collate and save software versions def topic_versions = channel.topic("versions") .distinct() .branch { entry -> @@ -219,9 +216,7 @@ workflow ASSEMBLYSUBMIT { newLine: true ) - // - // MODULE: MultiQC - // + // --------- MODULE: MultiQC ch_multiqc_files = ch_multiqc_files.mix(ch_collated_versions) def ch_summary_params = paramsSummaryMap(workflow, parameters_schema: "nextflow_schema.json") def ch_workflow_summary = channel.value(paramsSummaryMultiqc(ch_summary_params)) diff --git a/workflows/genomesubmit.nf b/workflows/genomesubmit.nf index 780a88e..d16de68 100644 --- a/workflows/genomesubmit.nf +++ b/workflows/genomesubmit.nf @@ -59,7 +59,7 @@ workflow GENOMESUBMIT { def ch_versions = channel.empty() def ch_multiqc_files = channel.empty() - // --------- Create genomes channel with proper metadata structure + // --------- Create genomes channel with proper metadata structure genome_fasta_and_reads = ch_samplesheet .map { row -> def meta = [ @@ -102,19 +102,17 @@ workflow GENOMESUBMIT { ) // --------- Genome coverage calculation - FASTA_VALIDATION.out.valid_fastas + branched_coverage_results = FASTA_VALIDATION.out.valid_fastas .branch { meta, _fasta -> genome_coverage_ref_input: meta.genome_coverage == null genome_coverage_present: true // Everything else goes here } - .set { branched_coverage_results } - branched_coverage_results.genome_coverage_ref_input.join(genome_reads) + coverm_input = branched_coverage_results.genome_coverage_ref_input.join(genome_reads) .multiMap { meta, fasta, fastq -> genome: [ meta, fasta ] raw_reads: [ meta, fastq ] } - .set { coverm_input } COVERM_GENOME ( coverm_input.raw_reads, @@ -138,12 +136,11 @@ workflow GENOMESUBMIT { .mix(branched_coverage_results.genome_coverage_present) // --------- For genomes without RNA_presence info, calculate rRNA and tRNA - fasta_updated_with_coverage + branched_rna_results = fasta_updated_with_coverage .branch { meta, _fasta -> rna_prediction_input: meta.RNA_presence == null rna_present: true // Everything else goes here } - .set { branched_rna_results } RNA_DETECTION ( branched_rna_results.rna_prediction_input, @@ -164,26 +161,16 @@ workflow GENOMESUBMIT { .mix(branched_rna_results.rna_present) // --------- Completeness and contamination calculation - fasta_updated_with_rna + branched_stats_results = fasta_updated_with_rna .branch { meta, _fasta -> genome_evaluation_input: meta.completeness == null || meta.contamination == null || meta.stats_generation_software == null evaluation_present: true // Everything else goes here } - .set { branched_stats_results } - - // build input structures for CheckM2 DB depending on what provided as input - def checkm2_db_input = checkm2_db - ? channel.of( [['id': 'CHECKM2_DB'], file(checkm2_db)] ) - : channel.empty() - - def checkm2_db_id_input = (!checkm2_db && checkm2_db_download_id) - ? channel.of( [['id': 'CHECKM2_DB_id'], checkm2_db_download_id] ) - : channel.empty() GENOME_EVALUATION ( branched_stats_results.genome_evaluation_input, - checkm2_db_input, - checkm2_db_id_input + checkm2_db, + checkm2_db_download_id ) // Create a value channel with the version string @@ -206,12 +193,11 @@ workflow GENOMESUBMIT { .mix(branched_stats_results.evaluation_present) // --------- Taxonomy - fasta_updated_with_stats + branched_taxonomy_results = fasta_updated_with_stats .branch { meta, _fasta -> genome_taxonomy_input: meta.NCBI_lineage == null taxonomy_present: true // Everything else goes here } - .set { branched_taxonomy_results } // Change extension for all files required taxonomy to .fasta because CATPACK requires suffix as input RENAME_FASTA_FOR_CATPACK ( @@ -258,10 +244,7 @@ workflow GENOMESUBMIT { 'true' // skip_header - we want to keep the header from the first file and skip it for the rest ) - // --------- Register study if accession not provided - if (!submission_study && !study_metadata) { - error("Either --submission_study or --study_metadata must be provided") - } + // --------- Register study if accession not provided via --submission_study def study_accession_ch if (submission_study) { study_accession_ch = channel.of(submission_study) @@ -312,7 +295,6 @@ workflow GENOMESUBMIT { } // --------- Upload data to ENA - SUBMIT ( ch_combined, test_upload, @@ -326,9 +308,7 @@ workflow GENOMESUBMIT { 'true' // skip_header - we want to keep the header from the first file and skip it for the rest ) - // - // Collate and save software versions - // + // --------- Collate and save software versions def topic_versions = channel.topic("versions") .distinct() .branch { entry -> @@ -355,10 +335,7 @@ workflow GENOMESUBMIT { newLine: true ) - - // - // MODULE: MultiQC - // + // --------- MODULE: MultiQC ch_multiqc_files = ch_multiqc_files.mix(ch_collated_versions) def ch_summary_params = paramsSummaryMap(workflow, parameters_schema: "nextflow_schema.json") def ch_workflow_summary = channel.value(paramsSummaryMultiqc(ch_summary_params)) diff --git a/workflows/readsubmit.nf b/workflows/readsubmit.nf index 547b466..8b90399 100644 --- a/workflows/readsubmit.nf +++ b/workflows/readsubmit.nf @@ -43,7 +43,7 @@ workflow READSUBMIT { def ch_versions = channel.empty() def ch_multiqc_files = channel.empty() - // Create reads channel with proper metadata structure + // --------- Create reads channel with proper metadata structure reads_ch = ch_samplesheet .map { row -> def (meta_in, sample_accession, fastq_1, fastq_2, @@ -72,9 +72,7 @@ workflow READSUBMIT { } } - if (!submission_study && !study_metadata) { - error("Either --submission_study or --study_metadata must be provided") - } + // --------- Register study if accession was not provided via --submission_study def study_accession_ch if (submission_study) { // Use provided study accession directly @@ -94,7 +92,7 @@ workflow READSUBMIT { } } - // Generate reads manifest files + // --------- Generate reads manifest files CREATE_READS_MANIFEST( reads_ch, study_accession_ch.first() @@ -106,6 +104,7 @@ workflow READSUBMIT { [meta, fastq, manifest] } + // --------- Upload data to ENA SUBMIT ( submission_input, test_upload, @@ -119,9 +118,7 @@ workflow READSUBMIT { true // skip_header - we want to keep the header from the first file and skip it for the rest ) - // - // Collate and save software versions - // + // --------- Collate and save software versions def topic_versions = channel.topic("versions") .distinct() .branch { entry -> @@ -148,9 +145,7 @@ workflow READSUBMIT { newLine: true ) - // - // MODULE: MultiQC - // + // --------- MODULE: MultiQC ch_multiqc_files = ch_multiqc_files.mix(ch_collated_versions) def ch_summary_params = paramsSummaryMap(workflow, parameters_schema: "nextflow_schema.json") def ch_workflow_summary = channel.value(paramsSummaryMultiqc(ch_summary_params)) From 1a31a0f6ca60740c5231fb4db8d0d7ea959a67fe Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Tue, 21 Jul 2026 20:56:35 +0100 Subject: [PATCH 29/47] rename sample to id in samplesheet and add fields to meta directly from schema --- README.md | 14 ++------- assets/schema_input_assembly.json | 19 ++++++----- assets/schema_input_genome.json | 52 ++++++++++++++++++++----------- assets/schema_input_reads.json | 33 +++++++++++++------- docs/usage.md | 27 ++++++---------- workflows/assemblysubmit.nf | 31 +++++------------- workflows/genomesubmit.nf | 31 +++--------------- workflows/readsubmit.nf | 26 +++------------- 8 files changed, 97 insertions(+), 136 deletions(-) diff --git a/README.md b/README.md index e3e201a..f7b381c 100644 --- a/README.md +++ b/README.md @@ -21,14 +21,6 @@ ## Introduction -**nf-core/seqsubmit** is a Nextflow pipeline for submitting sequence data to [ENA](https://www.ebi.ac.uk/ena/browser/home). -Currently, the pipeline supports four submission modes, each routed to a dedicated workflow and requiring its own input samplesheet structure: - -- `mags` for Metagenome Assembled Genomes (MAGs) submission with `GENOMESUBMIT` workflow -- `bins` for bins submission with `GENOMESUBMIT` workflow -- `metagenomic_assemblies` for assembly submission with `ASSEMBLYSUBMIT` workflow -- `reads` for raw sequencing reads submission with `READSUBMIT` workflow - ![seqsubmit workflow diagram](docs/images/seqsubmit_schema.png) **nf-core/seqsubmit** is a Nextflow pipeline for submitting sequence data to [ENA](https://www.ebi.ac.uk/ena/browser/home). @@ -68,7 +60,7 @@ Each workflow has its own samplesheet structure, prerequisites, and limitations Example: ```csv -sample,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,library_selection,library_strategy,insert_size,library_name,description +id,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,library_selection,library_strategy,insert_size,library_name,description illumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLUMINA,Illumina HiSeq 2000,GENOMIC,RANDOM,WGS,500,HiSeq_library_001,Illumina sequencing of sample XYZ ``` @@ -79,7 +71,7 @@ See the [`reads` mode section](docs/usage.md#samplesheet-input) of the usage doc Example: ```csv -sample,fasta,fastq_1,fastq_2,coverage,run_accession,assembler,assembler_version +id,fasta,fastq_1,fastq_2,coverage,run_accession,assembler,assembler_version assembly_1,data/contigs_1.fasta.gz,data/reads_1.fastq.gz,data/reads_2.fastq.gz,,ERR011322,SPAdes,3.15.5 assembly_2,data/contigs_2.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 ``` @@ -91,7 +83,7 @@ See the [`metagenomic_assemblies` mode section](docs/usage.md#samplesheet-input- Example: ```csv -sample,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage +id,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage lachnospira_eligens,data/bin_lachnospira_eligens.fa.gz,SRR24458089,,,spades_v3.15.5,metabat2_v2.6,default,CheckM2_v1.0.1,61.0,0.21,32.07,sediment metagenome,No,marine,cable_bacteria,marine_sediment,No,d__Bacteria;p__Proteobacteria;s__unclassified_Proteobacteria ``` diff --git a/assets/schema_input_assembly.json b/assets/schema_input_assembly.json index 6123501..78d1d60 100644 --- a/assets/schema_input_assembly.json +++ b/assets/schema_input_assembly.json @@ -7,10 +7,11 @@ "items": { "type": "object", "properties": { - "sample": { + "id": { "type": "string", "pattern": "^\\S+$", - "errorMessage": "Sample must be provided and cannot contain spaces", + "description": "Unique assembly identifier", + "errorMessage": "ID must be provided and cannot contain spaces", "meta": ["id"] }, "fasta": { @@ -65,28 +66,32 @@ } ], "errorMessage": "Coverage must be a positive number or empty", - "description": "Estimated value of assembly coverage" + "description": "Estimated value of assembly coverage", + "meta": ["coverage"] }, "run_accession": { "type": "string", "pattern": "^\\S+$", "errorMessage": "Accession must be provided and cannot contain spaces", - "description": "Accession of the run used to generate the assembly" + "description": "Accession of the run used to generate the assembly", + "meta": ["run_accession"] }, "assembler": { "type": "string", "pattern": "^\\S+$", "errorMessage": "Assembler must be provided and cannot contain spaces", - "description": "Name of the assembler software used to generate the assembly, e.g. MEGAHIT, SPAdes, metaSPAdes, Flye, etc" + "description": "Name of the assembler software used to generate the assembly, e.g. MEGAHIT, SPAdes, metaSPAdes, Flye, etc", + "meta": ["assembler"] }, "assembler_version": { "anyOf": [{ "type": "string" }, { "type": "number" }], "pattern": "^\\S+$", "errorMessage": "Assembler version must be provided and cannot contain spaces", - "description": "Version of the assembler software used to generate the assembly, e.g 1.0.0" + "description": "Version of the assembler software used to generate the assembly, e.g 1.0.0", + "meta": ["assembler_version"] } }, - "required": ["sample", "fasta", "run_accession", "assembler", "assembler_version"], + "required": ["id", "fasta", "run_accession", "assembler", "assembler_version"], "anyOf": [ { "properties": { diff --git a/assets/schema_input_genome.json b/assets/schema_input_genome.json index a603b2c..db21b28 100644 --- a/assets/schema_input_genome.json +++ b/assets/schema_input_genome.json @@ -7,10 +7,11 @@ "items": { "type": "object", "properties": { - "sample": { + "id": { "type": "string", "pattern": "^\\S+$", - "errorMessage": "Sample name must be provided and cannot contain spaces", + "description": "Unique mag/bin identifier", + "errorMessage": "ID must be provided and cannot contain spaces", "meta": ["id"] }, "fasta": { @@ -23,7 +24,8 @@ }, "accession": { "type": "string", - "description": "Run or assembly ENA accession" + "description": "Run or assembly ENA accession", + "meta": ["accession"] }, "fastq_1": { "anyOf": [ @@ -59,69 +61,83 @@ }, "assembly_software": { "type": "string", - "description": "Tool name and version that was used to assemble data, e.g. MEGAHIT_v1.0, SPAdes_v4.0.0, metaSPAdes_v3.15.0" + "description": "Tool name and version that was used to assemble data, e.g. MEGAHIT_v1.0, SPAdes_v4.0.0, metaSPAdes_v3.15.0", + "meta": ["assembly_software"] }, "binning_software": { "type": "string", - "description": "Tool name and version that was used to bin data, e.g. MetaBAT2, VAMB, MaxBin2, CONCOCT, SemiBin2, COMEBin, etc" + "description": "Tool name and version that was used to bin data, e.g. MetaBAT2, VAMB, MaxBin2, CONCOCT, SemiBin2, COMEBin, etc", + "meta": ["binning_software"] }, "binning_parameters": { "type": "string", - "description": "Arguments used to bin data different from default, e.g. -min_contig_length 1000" + "description": "Arguments used to bin data different from default, e.g. -min_contig_length 1000", + "meta": ["binning_parameters"] }, "stats_generation_software": { "type": "string", - "description": "Tool(s) used to estimate completeness and contamination" + "description": "Tool(s) used to estimate completeness and contamination", + "meta": ["stats_generation_software"] }, "completeness": { "type": "number", "description": "MAG/bin completeness score: the ratio of observed single-copy marker genes to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined", "exclusiveMinimum": 0, - "exclusiveMaximum": 100 + "exclusiveMaximum": 100, + "meta": ["completeness"] }, "contamination": { "type": "number", "description": "MAG/bin contamination score: the ratio of observed single-copy marker genes in ≥2 copies to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined", "exclusiveMinimum": 0, - "exclusiveMaximum": 100 + "exclusiveMaximum": 100, + "meta": ["contamination"] }, "genome_coverage": { "type": "number", - "description": "MAG/bin coverage value" + "description": "MAG/bin coverage value", + "meta": ["genome_coverage"] }, "metagenome": { "type": "string", - "description": "ENA accepted metagenome name. It needs to be listed in the taxonomy tree https://www.ebi.ac.uk/ena/browser/view/408169?show=tax-tree (you might need to press \"Tax tree - Show\" in the right most section of the page). Full list can also be found https://github.com/EBI-Metagenomics/genome_uploader/blob/main/genomeuploader/constants.py#L22" + "description": "ENA accepted metagenome name. It needs to be listed in the taxonomy tree https://www.ebi.ac.uk/ena/browser/view/408169?show=tax-tree (you might need to press \"Tax tree - Show\" in the right most section of the page). Full list can also be found https://github.com/EBI-Metagenomics/genome_uploader/blob/main/genomeuploader/constants.py#L22", + "meta": ["metagenome"] }, "co-assembly": { "type": "string", "description": "'Yes' if data was co-assembled, 'No' otherwise. Options: 'Yes' or 'No'", - "enum": ["Yes", "No"] + "enum": ["Yes", "No"], + "meta": ["co_assembly"] }, "broad_environment": { "type": "string", - "description": "Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428). Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS" + "description": "Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428). Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS", + "meta": ["broad_environment"] }, "local_environment": { "type": "string", - "description": "Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for \"broad-scale environmental context\". Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS" + "description": "Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for \"broad-scale environmental context\". Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS", + "meta": ["local_environment"] }, "environmental_medium": { "type": "string", - "description": "Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS" + "description": "Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS", + "meta": ["environmental_medium"] }, "RNA_presence": { "type": "string", "description": "Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: 'Yes' or 'No'.", - "enum": ["Yes", "No"] + "enum": ["Yes", "No"], + "meta": ["RNA_presence"] }, "NCBI_lineage": { "type": "string", - "description": "full NCBI lineage - format: x;y;z. For example, the lineage for E. coli can be: 'Bacteria;Pseudomonadati;Pseudomonadota;Gammaproteobacteria;Enterobacterales;Enterobacteriaceae;Escherichia' or '2;1224;1236;91347;543;561;562'. For more info check https://www.ncbi.nlm.nih.gov/datasets/docs/v2/data-processing/taxonomy-processing/taxonomy/" + "description": "full NCBI lineage - format: x;y;z. For example, the lineage for E. coli can be: 'Bacteria;Pseudomonadati;Pseudomonadota;Gammaproteobacteria;Enterobacterales;Enterobacteriaceae;Escherichia' or '2;1224;1236;91347;543;561;562'. For more info check https://www.ncbi.nlm.nih.gov/datasets/docs/v2/data-processing/taxonomy-processing/taxonomy/", + "meta": ["NCBI_lineage"] } }, "required": [ - "sample", + "id", "fasta", "accession", "assembly_software", diff --git a/assets/schema_input_reads.json b/assets/schema_input_reads.json index 25dfb31..5aa8887 100644 --- a/assets/schema_input_reads.json +++ b/assets/schema_input_reads.json @@ -7,10 +7,10 @@ "items": { "type": "object", "properties": { - "sample": { + "id": { "type": "string", "pattern": "^\\S+$", - "errorMessage": "Sample must be provided and cannot contain spaces", + "errorMessage": "ID must be provided and cannot contain spaces", "meta": ["id"], "description": "Unique experiment/run name" }, @@ -18,7 +18,8 @@ "type": "string", "pattern": "^\\S+$", "errorMessage": "Sample accession must be provided and cannot contain spaces", - "description": "ENA sample accession of the sample used to generate the reads" + "description": "ENA sample accession of the sample used to generate the reads", + "meta": ["sample_accession"] }, "fastq_1": { "type": "string", @@ -63,13 +64,15 @@ "ULTIMA", "VELA_DIAGNOSTICS" ], - "description": "Sequencing platform. Must be one of the ENA controlled vocabulary values listed in the enum." + "description": "Sequencing platform. Must be one of the ENA controlled vocabulary values listed in the enum.", + "meta": ["platform"] }, "instrument": { "type": "string", "pattern": "^[^\\n]+$", "errorMessage": "Instrument must be provided and cannot span multiple lines", - "description": "Sequencer model (e.g., 'Illumina HiSeq 2000', 'PacBio Sequel')" + "description": "Sequencer model (e.g., 'Illumina HiSeq 2000', 'PacBio Sequel')", + "meta": ["instrument"] }, "library_source": { "type": "string", @@ -84,7 +87,8 @@ "VIRAL RNA", "OTHER" ], - "description": "Library source. Must be one of the ENA controlled vocabulary values listed in the enum." + "description": "Library source. Must be one of the ENA controlled vocabulary values listed in the enum.", + "meta": ["library_source"] }, "library_selection": { "type": "string", @@ -121,7 +125,8 @@ "other", "unspecified" ], - "description": "Library selection. Must be one of the ENA controlled vocabulary values listed in the enum." + "description": "Library selection. Must be one of the ENA controlled vocabulary values listed in the enum.", + "meta": ["library_selection"] }, "library_strategy": { "type": "string", @@ -168,7 +173,8 @@ "Tethered Chromatin Conformation Capture", "OTHER" ], - "description": "Library strategy. Must be one of the ENA controlled vocabulary values listed in the enum." + "description": "Library strategy. Must be one of the ENA controlled vocabulary values listed in the enum.", + "meta": ["library_strategy"] }, "insert_size": { "anyOf": [ @@ -182,7 +188,8 @@ } ], "errorMessage": "Insert size must be a positive number or empty", - "description": "Fragment/insert size for paired-end reads (optional)" + "description": "Fragment/insert size for paired-end reads (optional)", + "meta": ["insert_size"] }, "library_name": { "anyOf": [ @@ -194,7 +201,8 @@ "maxLength": 0 } ], - "description": "Descriptive library name (optional)" + "description": "Descriptive library name (optional)", + "meta": ["library_name"] }, "description": { "anyOf": [ @@ -206,11 +214,12 @@ "maxLength": 0 } ], - "description": "Free-text description of the experiment (optional)" + "description": "Free-text description of the experiment (optional)", + "meta": ["description"] } }, "required": [ - "sample", + "id", "sample_accession", "fastq_1", "platform", diff --git a/docs/usage.md b/docs/usage.md index ea3a386..8b79848 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -156,17 +156,14 @@ You will need to create a samplesheet with information about the raw reads you w Example: ```csv title="samplesheet_reads.csv" -sample,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,library_selection,library_strategy,insert_size,library_name,description +id,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,library_selection,library_strategy,insert_size,library_name,description illumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLUMINA,Illumina HiSeq 2000,GENOMIC,RANDOM,WGS,500,HiSeq_library_001,Illumina sequencing of sample XYZ pacbio_run_001,SAMEA7654321,data/pacbio_reads.fastq.gz,,PACBIO_SMRT,PacBio Sequel,GENOMIC,RANDOM,WGS,,PacBio_library_002,Long-read sequencing ``` -> [!IMPORTANT] -> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. - | Column | Required | Description | -| ------------------- | -------- | --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | Yes | Unique identifier of this particular data entry. Used as an experiment name. | +|---------------------| -------- | --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `id` | Yes | Unique identifier of this particular data entry. Used as an experiment name. | | `sample_accession` | Yes | ENA sample accession (starting with SAMEA) of the sample used to generate raw reads. | | `fastq_1` | Yes | Path to forward reads in FASTQ format (optionally gzipped). | | `fastq_2` | No | Path to reverse reads for paired-end data. Leave empty for single-end reads. | @@ -217,17 +214,14 @@ You will need to create a samplesheet with information about the metagenomic ass Example: ```csv title="samplesheet_assembly.csv" -sample,fasta,fastq_1,fastq_2,coverage,run_accession,assembler,assembler_version +id,fasta,fastq_1,fastq_2,coverage,run_accession,assembler,assembler_version assembly_001,data/assembly_001.fasta.gz,data/assembly_001_R1.fastq.gz,data/assembly_001_R2.fastq.gz,,ERR011322,SPAdes,3.15.5 assembly_002,data/assembly_002.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 ``` -> [!IMPORTANT] -> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. - | Column | Required | Description | -| ------------------- | ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset. | +|---------------------| ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------- | +| `id` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset. | | `fasta` | Yes | Path to assembly contigs in FASTA format compressed with `gzip`. | | `fastq_1` | Conditional | Path to the read file in FASTQ format used to generate the metagenomic assembly. Required if `coverage` is not provided. | | `fastq_2` | No | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | @@ -299,16 +293,13 @@ You will need to create a samplesheet with information about the MAGs/bins you w Example: ```csv title="samplesheet_genomes.csv" -sample,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage +id,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default,CheckM2 1.0.1,92.81,1.09,66.04,sediment metagenome,No,marine,cable bacteria,marine sediment,No,d__Bacteria;p__Proteobacteria;s__ ``` -> [!IMPORTANT] -> **Samplesheet column requirements**: All columns shown in the example above must be present in your samplesheet, even if some values are empty. Columns must be in exactly the same order as shown. - | Column | Required | Description | -| --------------------------- | ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset. | +|-----------------------------| ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `id` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset. | | `fasta` | Yes | Path to MAG/bin contigs in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. | | `accession` | Yes | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | | `fastq_1` | Conditional | Path to the read file in FASTQ format used to generate the source metagenomic assembly. Required if `genome_coverage` is not provided. | diff --git a/workflows/assemblysubmit.nf b/workflows/assemblysubmit.nf index b209609..64b524e 100644 --- a/workflows/assemblysubmit.nf +++ b/workflows/assemblysubmit.nf @@ -50,37 +50,22 @@ workflow ASSEMBLYSUBMIT { // --------- Create assembly channel with proper metadata structure assembly_fasta = ch_samplesheet - .map { row -> - def meta = [ - id: row[0].id, - single_end: row[3] ? false : true, - coverage: row[4] ?: null, - run_accession: row[5], - assembler: row[6], - assembler_version: row[7] - ] - [meta, file(row[1])] + .map { meta, fasta, reads_1, reads_2 -> + def new_meta = meta + [single_end: reads_2 ? false : true] + [new_meta, fasta] } // --------- Create reads channel with proper metadata structure reads_fastq = ch_samplesheet .filter { row -> row[2] && row[2] != "" } // Check if fastq_1 exists and is not empty - .map { row -> - def meta = [ - id: row[0].id, - single_end: row[3] ? false : true, - coverage: row[4] ?: null, - run_accession: row[5], - assembler: row[6], - assembler_version: row[7] - ] - - if (row[3] && row[3] != "") { + .map { meta, fasta, reads_1, reads_2 -> + def new_meta = meta + [single_end: reads_2 ? false : true] + if (reads_2 && reads_2 != "") { // If paired end reads - [meta, [file(row[2]), file(row[3])]] + [new_meta, [file(reads_1), file(reads_2)]] } else { // If single end - [meta, file(row[2])] + [new_meta, file(reads_1)] } } diff --git a/workflows/genomesubmit.nf b/workflows/genomesubmit.nf index d16de68..098b037 100644 --- a/workflows/genomesubmit.nf +++ b/workflows/genomesubmit.nf @@ -61,35 +61,14 @@ workflow GENOMESUBMIT { // --------- Create genomes channel with proper metadata structure genome_fasta_and_reads = ch_samplesheet - .map { row -> - def meta = [ - id: row[0].id, - accession: row[2], - single_end: row[4] ? false : true, - assembly_software: row[5] ?: null, - binning_software: row[6] ?: null, - binning_parameters: row[7] ?: null, - stats_generation_software: row[8] ?: null, - completeness: row[9] ?: null, - contamination: row[10] ?: null, - genome_coverage: row[11] ?: null, - metagenome: row[12] ?: null, - co_assembly: row[13] ?: null, - broad_environment: row[14] ?: null, - local_environment: row[15] ?: null, - environmental_medium: row[16] ?: null, - RNA_presence: row[17] ?: null, - NCBI_lineage: row[18] ?: null - ] - def read1 = row[3] ? file(row[3]) : null - def read2 = row[4] ? file(row[4]) : null - - if (row[4] && row[4] != "") { + .map { meta, fasta, reads_1, reads_2 -> + def new_meta = meta + [single_end: reads_2 ? false : true] + if (reads_2 && reads_2 != "") { // If paired end reads - return [meta, file(row[1]), [read1, read2]] + return [new_meta, file(fasta), [reads_1, reads_2]] } else { // If single end - return [meta, file(row[1]), [read1]] + return [new_meta, file(fasta), [reads_1]] } } diff --git a/workflows/readsubmit.nf b/workflows/readsubmit.nf index 8b90399..614ae03 100644 --- a/workflows/readsubmit.nf +++ b/workflows/readsubmit.nf @@ -45,30 +45,14 @@ workflow READSUBMIT { // --------- Create reads channel with proper metadata structure reads_ch = ch_samplesheet - .map { row -> - def (meta_in, sample_accession, fastq_1, fastq_2, - platform, instrument, library_source, library_selection, library_strategy, - insert_size, library_name, description) = row - def meta = [ - id: meta_in.id, - sample_accession: sample_accession, - single_end: fastq_2 ? false : true, - platform: platform, - instrument: instrument, - library_source: library_source, - library_selection: library_selection, - library_strategy: library_strategy, - insert_size: insert_size ?: null, - library_name: library_name ?: null, - description: description ?: null - ] - - if (fastq_2 && fastq_2 != "") { + .map { meta, reads_1, reads_2 -> + def new_meta = meta + [single_end: reads_2 ? false : true] + if (reads_2 && reads_2 != "") { // If paired end reads - [meta, [file(fastq_1), file(fastq_2)]] + [new_meta, [file(reads_1), file(reads_2)]] } else { // If single end - [meta, file(fastq_1)] + [new_meta, file(reads_1)] } } From f1228e7a950f51161faae2707efdffb2d0ac3972 Mon Sep 17 00:00:00 2001 From: Martin Beracochea Date: Wed, 22 Jul 2026 09:36:13 +0100 Subject: [PATCH 30/47] Fix prettier issues with docs/usage.md --- docs/usage.md | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index 8b79848..4716cc8 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -162,7 +162,7 @@ pacbio_run_001,SAMEA7654321,data/pacbio_reads.fastq.gz,,PACBIO_SMRT,PacBio Seque ``` | Column | Required | Description | -|---------------------| -------- | --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| ------------------- | -------- | --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | | `id` | Yes | Unique identifier of this particular data entry. Used as an experiment name. | | `sample_accession` | Yes | ENA sample accession (starting with SAMEA) of the sample used to generate raw reads. | | `fastq_1` | Yes | Path to forward reads in FASTQ format (optionally gzipped). | @@ -220,7 +220,7 @@ assembly_002,data/assembly_002.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 ``` | Column | Required | Description | -|---------------------| ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------- | +| ------------------- | ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------- | | `id` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset. | | `fasta` | Yes | Path to assembly contigs in FASTA format compressed with `gzip`. | | `fastq_1` | Conditional | Path to the read file in FASTQ format used to generate the metagenomic assembly. Required if `coverage` is not provided. | @@ -298,7 +298,7 @@ mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default, ``` | Column | Required | Description | -|-----------------------------| ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| --------------------------- | ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | | `id` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset. | | `fasta` | Yes | Path to MAG/bin contigs in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. | | `accession` | Yes | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | From 0a6e44b72b0419245813a9b3ff4f3f75f935fcc4 Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Wed, 22 Jul 2026 11:12:39 +0100 Subject: [PATCH 31/47] tidy up .map after review --- assets/schema_input_genome.json | 2 +- assets/schema_input_reads.json | 2 +- workflows/assemblysubmit.nf | 6 +++--- workflows/genomesubmit.nf | 4 ++-- workflows/readsubmit.nf | 4 ++-- 5 files changed, 9 insertions(+), 9 deletions(-) diff --git a/assets/schema_input_genome.json b/assets/schema_input_genome.json index db21b28..1ae38f1 100644 --- a/assets/schema_input_genome.json +++ b/assets/schema_input_genome.json @@ -132,7 +132,7 @@ }, "NCBI_lineage": { "type": "string", - "description": "full NCBI lineage - format: x;y;z. For example, the lineage for E. coli can be: 'Bacteria;Pseudomonadati;Pseudomonadota;Gammaproteobacteria;Enterobacterales;Enterobacteriaceae;Escherichia' or '2;1224;1236;91347;543;561;562'. For more info check https://www.ncbi.nlm.nih.gov/datasets/docs/v2/data-processing/taxonomy-processing/taxonomy/", + "description": "Full NCBI lineage - format: x;y;z. For example, the lineage for E. coli can be: 'Bacteria;Pseudomonadati;Pseudomonadota;Gammaproteobacteria;Enterobacterales;Enterobacteriaceae;Escherichia' or '2;1224;1236;91347;543;561;562'. For more info check https://www.ncbi.nlm.nih.gov/datasets/docs/v2/data-processing/taxonomy-processing/taxonomy/", "meta": ["NCBI_lineage"] } }, diff --git a/assets/schema_input_reads.json b/assets/schema_input_reads.json index 5aa8887..d0c0df0 100644 --- a/assets/schema_input_reads.json +++ b/assets/schema_input_reads.json @@ -125,7 +125,7 @@ "other", "unspecified" ], - "description": "Library selection. Must be one of the ENA controlled vocabulary values listed in the enum.", + "description": "Library selection. Must be one of the allowed ENA controlled vocabulary values (e.g. PCR, ChIP-Seq, other, etc).", "meta": ["library_selection"] }, "library_strategy": { diff --git a/workflows/assemblysubmit.nf b/workflows/assemblysubmit.nf index 64b524e..f472653 100644 --- a/workflows/assemblysubmit.nf +++ b/workflows/assemblysubmit.nf @@ -51,7 +51,7 @@ workflow ASSEMBLYSUBMIT { // --------- Create assembly channel with proper metadata structure assembly_fasta = ch_samplesheet .map { meta, fasta, reads_1, reads_2 -> - def new_meta = meta + [single_end: reads_2 ? false : true] + def new_meta = meta + [single_end: reads_2 == null] [new_meta, fasta] } @@ -59,8 +59,8 @@ workflow ASSEMBLYSUBMIT { reads_fastq = ch_samplesheet .filter { row -> row[2] && row[2] != "" } // Check if fastq_1 exists and is not empty .map { meta, fasta, reads_1, reads_2 -> - def new_meta = meta + [single_end: reads_2 ? false : true] - if (reads_2 && reads_2 != "") { + def new_meta = meta + [single_end: reads_2 == null] + if ( !new_meta.single_end ) { // If paired end reads [new_meta, [file(reads_1), file(reads_2)]] } else { diff --git a/workflows/genomesubmit.nf b/workflows/genomesubmit.nf index 098b037..6aef14d 100644 --- a/workflows/genomesubmit.nf +++ b/workflows/genomesubmit.nf @@ -62,8 +62,8 @@ workflow GENOMESUBMIT { // --------- Create genomes channel with proper metadata structure genome_fasta_and_reads = ch_samplesheet .map { meta, fasta, reads_1, reads_2 -> - def new_meta = meta + [single_end: reads_2 ? false : true] - if (reads_2 && reads_2 != "") { + def new_meta = meta + [single_end: reads_2 == null] + if ( !new_meta.single_end ) { // If paired end reads return [new_meta, file(fasta), [reads_1, reads_2]] } else { diff --git a/workflows/readsubmit.nf b/workflows/readsubmit.nf index 614ae03..45c375e 100644 --- a/workflows/readsubmit.nf +++ b/workflows/readsubmit.nf @@ -46,8 +46,8 @@ workflow READSUBMIT { // --------- Create reads channel with proper metadata structure reads_ch = ch_samplesheet .map { meta, reads_1, reads_2 -> - def new_meta = meta + [single_end: reads_2 ? false : true] - if (reads_2 && reads_2 != "") { + def new_meta = meta + [single_end: reads_2 == null] + if ( !new_meta.single_end ) { // If paired end reads [new_meta, [file(reads_1), file(reads_2)]] } else { From 5b262bf2a18e1bdaa2aee88c4339dbf023146dbc Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Wed, 22 Jul 2026 12:44:33 +0100 Subject: [PATCH 32/47] add null values as defaults for conditional args --- assets/schema_input_assembly.json | 13 +++---------- assets/schema_input_genome.json | 6 ++++++ conf/containers_docker_arm64.config | 1 + modules/local/genome_upload/main.nf | 4 ++-- subworkflows/local/genome_evaluation/main.nf | 11 +++++++---- workflows/assemblysubmit.nf | 2 +- workflows/genomesubmit.nf | 8 ++++---- 7 files changed, 24 insertions(+), 21 deletions(-) diff --git a/assets/schema_input_assembly.json b/assets/schema_input_assembly.json index 78d1d60..5e9108f 100644 --- a/assets/schema_input_assembly.json +++ b/assets/schema_input_assembly.json @@ -55,16 +55,9 @@ "description": "Reverse reads FASTQ file if paired-end. Leave empty for single-end reads" }, "coverage": { - "anyOf": [ - { - "type": "number", - "minimum": 0 - }, - { - "type": "string", - "maxLength": 0 - } - ], + "type": "number", + "default": null, + "minimum": 0, "errorMessage": "Coverage must be a positive number or empty", "description": "Estimated value of assembly coverage", "meta": ["coverage"] diff --git a/assets/schema_input_genome.json b/assets/schema_input_genome.json index 1ae38f1..6678a63 100644 --- a/assets/schema_input_genome.json +++ b/assets/schema_input_genome.json @@ -76,11 +76,13 @@ }, "stats_generation_software": { "type": "string", + "default": null, "description": "Tool(s) used to estimate completeness and contamination", "meta": ["stats_generation_software"] }, "completeness": { "type": "number", + "default": null, "description": "MAG/bin completeness score: the ratio of observed single-copy marker genes to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined", "exclusiveMinimum": 0, "exclusiveMaximum": 100, @@ -88,6 +90,7 @@ }, "contamination": { "type": "number", + "default": null, "description": "MAG/bin contamination score: the ratio of observed single-copy marker genes in ≥2 copies to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined", "exclusiveMinimum": 0, "exclusiveMaximum": 100, @@ -95,6 +98,7 @@ }, "genome_coverage": { "type": "number", + "default": null, "description": "MAG/bin coverage value", "meta": ["genome_coverage"] }, @@ -126,12 +130,14 @@ }, "RNA_presence": { "type": "string", + "default": null, "description": "Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: 'Yes' or 'No'.", "enum": ["Yes", "No"], "meta": ["RNA_presence"] }, "NCBI_lineage": { "type": "string", + "default": null, "description": "Full NCBI lineage - format: x;y;z. For example, the lineage for E. coli can be: 'Bacteria;Pseudomonadati;Pseudomonadota;Gammaproteobacteria;Enterobacterales;Enterobacteriaceae;Escherichia' or '2;1224;1236;91347;543;561;562'. For more info check https://www.ncbi.nlm.nih.gov/datasets/docs/v2/data-processing/taxonomy-processing/taxonomy/", "meta": ["NCBI_lineage"] } diff --git a/conf/containers_docker_arm64.config b/conf/containers_docker_arm64.config index 369f743..11a7ab9 100644 --- a/conf/containers_docker_arm64.config +++ b/conf/containers_docker_arm64.config @@ -1 +1,2 @@ +process { withName: 'FALINT' { container = 'community.wave.seqera.io/library/fa-lint:1.2.0--9dec293ee366e2bd' } } process { withName: 'MULTIQC' { container = 'community.wave.seqera.io/library/multiqc:1.35--5c84a5000a226ab5' } } diff --git a/modules/local/genome_upload/main.nf b/modules/local/genome_upload/main.nf index 095d9ca..a0abb62 100644 --- a/modules/local/genome_upload/main.nf +++ b/modules/local/genome_upload/main.nf @@ -4,8 +4,8 @@ process GENOME_UPLOAD { conda "${moduleDir}/environment.yml" container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/genome-uploader:3.0.1--pyhdfd78af_0': - 'quay.io/biocontainers/genome-uploader:3.0.1--pyhdfd78af_0' }" + 'https://depot.galaxyproject.org/singularity/genome-uploader:3.0.4--pyhdfd78af_0': + 'quay.io/biocontainers/genome-uploader:3.0.4--pyhdfd78af_0' }" input: path(mags) // required for validation diff --git a/subworkflows/local/genome_evaluation/main.nf b/subworkflows/local/genome_evaluation/main.nf index 4c477ee..e6862e1 100644 --- a/subworkflows/local/genome_evaluation/main.nf +++ b/subworkflows/local/genome_evaluation/main.nf @@ -29,13 +29,16 @@ workflow GENOME_EVALUATION { // // Database preparation // + ch_do_download = ch_fasta + .first() + .map { _meta_fasta -> ch_checkm2_db_download_id } // Download and prepare db from scratch if no pre-built db provided if (ch_checkm2_db) { - ch_checkm2_db = channel.value([[id: 'checkm2_db'], file(params.checkm2_db, checkIfExists: true)]) + ch_checkm2_db_ready = channel.value([[id: 'checkm2_db'], file(params.checkm2_db, checkIfExists: true)]) } else { - CHECKM2_DATABASEDOWNLOAD(ch_checkm2_db_download_id) - ch_checkm2_db = CHECKM2_DATABASEDOWNLOAD.out.database + CHECKM2_DATABASEDOWNLOAD(ch_do_download) + ch_checkm2_db_ready = CHECKM2_DATABASEDOWNLOAD.out.database } // @@ -44,7 +47,7 @@ workflow GENOME_EVALUATION { CHECKM2_PREDICT( ch_fasta, - ch_checkm2_db, + ch_checkm2_db_ready, ) emit: diff --git a/workflows/assemblysubmit.nf b/workflows/assemblysubmit.nf index f472653..401a32f 100644 --- a/workflows/assemblysubmit.nf +++ b/workflows/assemblysubmit.nf @@ -77,7 +77,7 @@ workflow ASSEMBLYSUBMIT { // --------- Assembly coverage calculation // For assemblies without coverage, calculate coverage with CoverM coverm_input = FASTA_VALIDATION.out.valid_fastas - .filter { meta, _fasta -> meta.coverage == null } + .filter { meta, _fasta -> !(meta.coverage) } .join(reads_fastq) .multiMap { meta, fasta, fastq -> assembly: [ meta, fasta ] diff --git a/workflows/genomesubmit.nf b/workflows/genomesubmit.nf index 6aef14d..be4cec9 100644 --- a/workflows/genomesubmit.nf +++ b/workflows/genomesubmit.nf @@ -83,7 +83,7 @@ workflow GENOMESUBMIT { // --------- Genome coverage calculation branched_coverage_results = FASTA_VALIDATION.out.valid_fastas .branch { meta, _fasta -> - genome_coverage_ref_input: meta.genome_coverage == null + genome_coverage_ref_input: !(meta.genome_coverage) genome_coverage_present: true // Everything else goes here } @@ -117,7 +117,7 @@ workflow GENOMESUBMIT { // --------- For genomes without RNA_presence info, calculate rRNA and tRNA branched_rna_results = fasta_updated_with_coverage .branch { meta, _fasta -> - rna_prediction_input: meta.RNA_presence == null + rna_prediction_input: !(meta.RNA_presence) rna_present: true // Everything else goes here } @@ -142,7 +142,7 @@ workflow GENOMESUBMIT { // --------- Completeness and contamination calculation branched_stats_results = fasta_updated_with_rna .branch { meta, _fasta -> - genome_evaluation_input: meta.completeness == null || meta.contamination == null || meta.stats_generation_software == null + genome_evaluation_input: !(meta.completeness) || !(meta.contamination) || !(meta.stats_generation_software) evaluation_present: true // Everything else goes here } @@ -174,7 +174,7 @@ workflow GENOMESUBMIT { // --------- Taxonomy branched_taxonomy_results = fasta_updated_with_stats .branch { meta, _fasta -> - genome_taxonomy_input: meta.NCBI_lineage == null + genome_taxonomy_input: !(meta.NCBI_lineage) taxonomy_present: true // Everything else goes here } From 54acdc989c1e1f67ac0bafae51f1b33afb7ec1fd Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Wed, 22 Jul 2026 12:58:00 +0100 Subject: [PATCH 33/47] update genome_uploader and fix regexp in multiqc --- assets/multiqc_config.yml | 6 +++--- tests/default.nf.test.snap | 2 +- tests/mag_complete_metadata.nf.test.snap | 2 +- tests/mag_no_coverage_paired_reads.nf.test.snap | 2 +- tests/mag_no_coverage_single_reads.nf.test.snap | 2 +- tests/mag_no_rna_presence.nf.test.snap | 2 +- tests/mag_no_study_complete_metadata.nf.test.snap | 2 +- tests/mag_no_taxonomy.nf.test.snap | 2 +- 8 files changed, 10 insertions(+), 10 deletions(-) diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index eb16eaf..9337e91 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -28,11 +28,11 @@ sp: genome_metadata: fn_re: ".*genomes_metadata.tsv$" sample_registration: - fn_re: '.*registered\.*\.tsv$' + fn_re: ".*registered*.tsv$" submission_results_genomes: - fn_re: 'genomes_accessions\.tsv$' + fn_re: "genomes_accessions.tsv$" submission_results_assemblies: - fn_re: 'assemblies_accessions\.tsv$' + fn_re: "assemblies_accessions.tsv$" # Custom data configuration custom_data: diff --git a/tests/default.nf.test.snap b/tests/default.nf.test.snap index 9968b79..4aadca3 100644 --- a/tests/default.nf.test.snap +++ b/tests/default.nf.test.snap @@ -35,7 +35,7 @@ "bash": "5.2.37(1)-release" }, "CREATE_MANIFESTS": { - "genome_uploader": "3.0.1" + "genome_uploader": "3.0.4" }, "FALINT": { "falint": "1.2.0" diff --git a/tests/mag_complete_metadata.nf.test.snap b/tests/mag_complete_metadata.nf.test.snap index 7c7f539..5495759 100644 --- a/tests/mag_complete_metadata.nf.test.snap +++ b/tests/mag_complete_metadata.nf.test.snap @@ -17,7 +17,7 @@ "bash": "5.2.37(1)-release" }, "CREATE_MANIFESTS": { - "genome_uploader": "3.0.1" + "genome_uploader": "3.0.4" }, "FALINT": { "falint": "1.2.0" diff --git a/tests/mag_no_coverage_paired_reads.nf.test.snap b/tests/mag_no_coverage_paired_reads.nf.test.snap index 894e756..0725d60 100644 --- a/tests/mag_no_coverage_paired_reads.nf.test.snap +++ b/tests/mag_no_coverage_paired_reads.nf.test.snap @@ -20,7 +20,7 @@ "bash": "5.2.37(1)-release" }, "CREATE_MANIFESTS": { - "genome_uploader": "3.0.1" + "genome_uploader": "3.0.4" }, "FALINT": { "falint": "1.2.0" diff --git a/tests/mag_no_coverage_single_reads.nf.test.snap b/tests/mag_no_coverage_single_reads.nf.test.snap index 767748f..3a43261 100644 --- a/tests/mag_no_coverage_single_reads.nf.test.snap +++ b/tests/mag_no_coverage_single_reads.nf.test.snap @@ -20,7 +20,7 @@ "bash": "5.2.37(1)-release" }, "CREATE_MANIFESTS": { - "genome_uploader": "3.0.1" + "genome_uploader": "3.0.4" }, "FALINT": { "falint": "1.2.0" diff --git a/tests/mag_no_rna_presence.nf.test.snap b/tests/mag_no_rna_presence.nf.test.snap index 8a7d21c..5541956 100644 --- a/tests/mag_no_rna_presence.nf.test.snap +++ b/tests/mag_no_rna_presence.nf.test.snap @@ -23,7 +23,7 @@ "bash": "5.2.37(1)-release" }, "CREATE_MANIFESTS": { - "genome_uploader": "3.0.1" + "genome_uploader": "3.0.4" }, "FALINT": { "falint": "1.2.0" diff --git a/tests/mag_no_study_complete_metadata.nf.test.snap b/tests/mag_no_study_complete_metadata.nf.test.snap index 1aea1e8..b838f6b 100644 --- a/tests/mag_no_study_complete_metadata.nf.test.snap +++ b/tests/mag_no_study_complete_metadata.nf.test.snap @@ -17,7 +17,7 @@ "bash": "5.2.37(1)-release" }, "CREATE_MANIFESTS": { - "genome_uploader": "3.0.1" + "genome_uploader": "3.0.4" }, "FALINT": { "falint": "1.2.0" diff --git a/tests/mag_no_taxonomy.nf.test.snap b/tests/mag_no_taxonomy.nf.test.snap index 1563d4b..0aa4bc4 100644 --- a/tests/mag_no_taxonomy.nf.test.snap +++ b/tests/mag_no_taxonomy.nf.test.snap @@ -26,7 +26,7 @@ "bash": "5.2.37(1)-release" }, "CREATE_MANIFESTS": { - "genome_uploader": "3.0.1" + "genome_uploader": "3.0.4" }, "FALINT": { "falint": "1.2.0" From 648ca9e1d36594f771a16dd29b3c1f55aeb43f45 Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Wed, 22 Jul 2026 15:12:41 +0100 Subject: [PATCH 34/47] test multiqc config old version from 1.0.0 branch --- assets/multiqc_config.yml | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index 9337e91..5df001f 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -28,11 +28,11 @@ sp: genome_metadata: fn_re: ".*genomes_metadata.tsv$" sample_registration: - fn_re: ".*registered*.tsv$" + fn_re: '.*registered.*\.tsv$' submission_results_genomes: - fn_re: "genomes_accessions.tsv$" + fn_re: 'genomes_accessions\.tsv$' submission_results_assemblies: - fn_re: "assemblies_accessions.tsv$" + fn_re: 'assemblies_accessions\.tsv$' # Custom data configuration custom_data: From edd594ae1db5adb77baeb574c3c001e1a40a7e29 Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Wed, 22 Jul 2026 17:47:47 +0100 Subject: [PATCH 35/47] change co-assembly and rna_presence to boolean --- README.md | 2 +- assets/schema_input_assembly.json | 13 ++- assets/schema_input_genome.json | 58 ++++++++++--- assets/schema_input_reads.json | 1 + bin/count_rna.py | 2 +- docs/methods.md | 6 +- docs/usage.md | 6 +- modules/local/count_rna/tests/main.nf.test | 2 +- .../local/count_rna/tests/main.nf.test.snap | 4 +- .../local/create_genome_metadata_tsv/main.nf | 4 +- .../tests/main.nf.test | 47 ++++++++++- .../tests/main.nf.test.snap | 84 ++++++++++++++++--- .../tests/main.nf.test | 11 +-- .../tests/main.nf.test.snap | 16 ++-- workflows/assemblysubmit.nf | 15 ++-- workflows/genomesubmit.nf | 17 ++-- workflows/readsubmit.nf | 6 +- 17 files changed, 212 insertions(+), 82 deletions(-) diff --git a/README.md b/README.md index f7b381c..f94503b 100644 --- a/README.md +++ b/README.md @@ -84,7 +84,7 @@ Example: ```csv id,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage -lachnospira_eligens,data/bin_lachnospira_eligens.fa.gz,SRR24458089,,,spades_v3.15.5,metabat2_v2.6,default,CheckM2_v1.0.1,61.0,0.21,32.07,sediment metagenome,No,marine,cable_bacteria,marine_sediment,No,d__Bacteria;p__Proteobacteria;s__unclassified_Proteobacteria +lachnospira_eligens,data/bin_lachnospira_eligens.fa.gz,SRR24458089,,,spades_v3.15.5,metabat2_v2.6,default,CheckM2_v1.0.1,61.0,0.21,32.07,sediment metagenome,false,marine,cable_bacteria,marine_sediment,false,d__Bacteria;p__Proteobacteria;s__unclassified_Proteobacteria ``` See the [`mags` and `bins` modes section](docs/usage.md#samplesheet-input-2) of the usage documentation for the full list of required and optional columns. diff --git a/assets/schema_input_assembly.json b/assets/schema_input_assembly.json index 5e9108f..c35a887 100644 --- a/assets/schema_input_assembly.json +++ b/assets/schema_input_assembly.json @@ -51,13 +51,22 @@ "maxLength": 0 } ], + "default": null, "errorMessage": "FASTQ file for reverse reads must have extension '.fq' or '.fastq' (optionally gzipped)", "description": "Reverse reads FASTQ file if paired-end. Leave empty for single-end reads" }, "coverage": { - "type": "number", + "anyOf": [ + { + "type": "number", + "minimum": 0 + }, + { + "type": "string", + "maxLength": 0 + } + ], "default": null, - "minimum": 0, "errorMessage": "Coverage must be a positive number or empty", "description": "Estimated value of assembly coverage", "meta": ["coverage"] diff --git a/assets/schema_input_genome.json b/assets/schema_input_genome.json index 6678a63..393211d 100644 --- a/assets/schema_input_genome.json +++ b/assets/schema_input_genome.json @@ -56,6 +56,7 @@ "maxLength": 0 } ], + "default": null, "errorMessage": "FASTQ file for reverse reads must have extension '.fq' or '.fastq' (optionally gzipped)", "description": "Reverse reads FASTQ file if paired-end. Leave empty for single-end reads" }, @@ -81,23 +82,48 @@ "meta": ["stats_generation_software"] }, "completeness": { - "type": "number", + "anyOf": [ + { + "type": "number", + "exclusiveMinimum": 0, + "exclusiveMaximum": 100 + }, + { + "type": "string", + "maxLength": 0 + } + ], "default": null, "description": "MAG/bin completeness score: the ratio of observed single-copy marker genes to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined", - "exclusiveMinimum": 0, - "exclusiveMaximum": 100, "meta": ["completeness"] }, "contamination": { - "type": "number", + "anyOf": [ + { + "type": "number", + "exclusiveMinimum": 0, + "exclusiveMaximum": 100 + }, + { + "type": "string", + "maxLength": 0 + } + ], "default": null, "description": "MAG/bin contamination score: the ratio of observed single-copy marker genes in ≥2 copies to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined", - "exclusiveMinimum": 0, - "exclusiveMaximum": 100, "meta": ["contamination"] }, "genome_coverage": { - "type": "number", + "anyOf": [ + { + "type": "number", + "minimum": 0 + }, + { + "type": "string", + "maxLength": 0 + } + ], "default": null, "description": "MAG/bin coverage value", "meta": ["genome_coverage"] @@ -108,9 +134,8 @@ "meta": ["metagenome"] }, "co-assembly": { - "type": "string", - "description": "'Yes' if data was co-assembled, 'No' otherwise. Options: 'Yes' or 'No'", - "enum": ["Yes", "No"], + "type": "boolean", + "description": "'true' if data was co-assembled, 'false' otherwise", "meta": ["co_assembly"] }, "broad_environment": { @@ -129,10 +154,17 @@ "meta": ["environmental_medium"] }, "RNA_presence": { - "type": "string", + "anyOf": [ + { + "type": "boolean" + }, + { + "type": "string", + "maxLength": 0 + } + ], "default": null, - "description": "Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: 'Yes' or 'No'.", - "enum": ["Yes", "No"], + "description": "Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: true/false", "meta": ["RNA_presence"] }, "NCBI_lineage": { diff --git a/assets/schema_input_reads.json b/assets/schema_input_reads.json index d0c0df0..e39c89e 100644 --- a/assets/schema_input_reads.json +++ b/assets/schema_input_reads.json @@ -42,6 +42,7 @@ "maxLength": 0 } ], + "default": null, "errorMessage": "FASTQ file for reverse reads must have extension '.fq' or '.fastq' (optionally gzipped)", "description": "Reverse reads FASTQ file if paired-end. Leave empty for single-end reads" }, diff --git a/bin/count_rna.py b/bin/count_rna.py index a514d37..8f34011 100755 --- a/bin/count_rna.py +++ b/bin/count_rna.py @@ -151,7 +151,7 @@ def main(): logging.info(f"Found {rrna_count} rRNA subunit types in total") # rrna_count should be 3 to make sure all 3 rRNA genes are present - decision = "Yes" if trna_count >= args.trna_limit and rrna_count == 3 else "No" + decision = "True" if trna_count >= args.trna_limit and rrna_count == 3 else "False" logging.info(f"RNA presented: {decision}") with open(args.output, 'w') as file_out: diff --git a/docs/methods.md b/docs/methods.md index 67e5488..567e19a 100644 --- a/docs/methods.md +++ b/docs/methods.md @@ -108,10 +108,10 @@ The total number of predicted tRNAs is compared against `params.trna_limit`. The The final decision stored in `RNA_presence` is: -- `Yes` when at least 18 tRNAs are detected and all three required rRNA subunits pass the recovery threshold -- `No` otherwise +- `True` when at least 18 tRNAs are detected and all three required rRNA subunits pass the recovery threshold +- `False` otherwise -The final `Yes`/`No` decision is then merged back into the submission metadata. +The final `True`/`False` decision is then merged back into the submission metadata. ### Genome coverage calculation diff --git a/docs/usage.md b/docs/usage.md index 4716cc8..353a906 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -294,7 +294,7 @@ Example: ```csv title="samplesheet_genomes.csv" id,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage -mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default,CheckM2 1.0.1,92.81,1.09,66.04,sediment metagenome,No,marine,cable bacteria,marine sediment,No,d__Bacteria;p__Proteobacteria;s__ +mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default,CheckM2 1.0.1,92.81,1.09,66.04,sediment metagenome,false,marine,cable bacteria,marine sediment,false,d__Bacteria;p__Proteobacteria;s__ ``` | Column | Required | Description | @@ -312,11 +312,11 @@ mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default, | `contamination` | No | Genome contamination value. | | `genome_coverage` | Conditional | Estimated average sequencing depth across the genome. If the value is missing, it is computed automatically during pipeline execution when reads are provided. | | `metagenome` | Yes | Registered metagenome taxonomic identifier or name that matches an existing ENA taxonomy entry. For more details see https://ena-docs.readthedocs.io/en/latest/faq/taxonomy.html | -| `co-assembly` | Yes | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: Yes or No. | +| `co-assembly` | Yes | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: true or false. | | `broad_environment` | Yes | Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428) | | `local_environment` | Yes | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". | | `environmental_medium` | Yes | Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). | -| `RNA_presence` | No | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: Yes or No. | +| `RNA_presence` | No | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: true or false. | | `NCBI_lineage` | No | NCBI taxonomy lineage of the genome. Can be composted of either numerical IDs or official NCBI taxon names separated by ";". | > [!NOTE] diff --git a/modules/local/count_rna/tests/main.nf.test b/modules/local/count_rna/tests/main.nf.test index d621839..657d437 100644 --- a/modules/local/count_rna/tests/main.nf.test +++ b/modules/local/count_rna/tests/main.nf.test @@ -31,7 +31,7 @@ nextflow_process { { assert process.out.rna_decision.size() == 1 }, { def rnaDecisionContent = path(process.out.rna_decision[0][1]).text - assert rnaDecisionContent.contains("Yes") + assert rnaDecisionContent.contains("True") } ) } diff --git a/modules/local/count_rna/tests/main.nf.test.snap b/modules/local/count_rna/tests/main.nf.test.snap index 46ab55a..047809f 100644 --- a/modules/local/count_rna/tests/main.nf.test.snap +++ b/modules/local/count_rna/tests/main.nf.test.snap @@ -6,11 +6,11 @@ { "id": "test" }, - "test_rna_decision.tsv:md5,8162813732f3a353edb6fc3e37e950e1" + "test_rna_decision.tsv:md5,b760b5d61d0c140d55e97a2fd6c25471" ] ] ], - "timestamp": "2026-06-25T21:38:21.745168", + "timestamp": "2026-07-22T17:43:41.374272", "meta": { "nf-test": "0.9.5", "nextflow": "26.04.0" diff --git a/modules/local/create_genome_metadata_tsv/main.nf b/modules/local/create_genome_metadata_tsv/main.nf index e78a7af..d3313f9 100644 --- a/modules/local/create_genome_metadata_tsv/main.nf +++ b/modules/local/create_genome_metadata_tsv/main.nf @@ -18,8 +18,8 @@ process CREATE_GENOME_METADATA_TSV { task.ext.when == null || task.ext.when script: - def co_assembly_value = meta.co_assembly == 'Yes' ? 'True' : 'False' - def rna_presence_value = meta.RNA_presence == 'Yes' ? 'True' : 'False' + def co_assembly_value = meta.co_assembly ? 'True' : 'False' + def rna_presence_value = meta.RNA_presence ? 'True' : 'False' def header = [ 'genome_name', 'genome_path', diff --git a/modules/local/create_genome_metadata_tsv/tests/main.nf.test b/modules/local/create_genome_metadata_tsv/tests/main.nf.test index eab3015..72516f5 100644 --- a/modules/local/create_genome_metadata_tsv/tests/main.nf.test +++ b/modules/local/create_genome_metadata_tsv/tests/main.nf.test @@ -24,11 +24,50 @@ nextflow_process { contamination: '1.2', genome_coverage: '42.7', metagenome: 'marine metagenome', - co_assembly: 'No', + co_assembly: false, broad_environment: 'marine biome', local_environment: 'coastal water', environmental_medium: 'seawater', - RNA_presence: 'Yes', + RNA_presence: true, + NCBI_lineage: 'Bacteria;Proteobacteria;Gammaproteobacteria' + ], + file('https://raw.githubusercontent.com/nf-core/test-datasets/viralrecon/genome/NC_045512.2/GCF_009858895.2_ASM985889v3_genomic.200409.fna.gz') + ] + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert snapshot(process.out).match() } + ) + } + + } + + test("test_create_genome_metadata_tsv_bool_reversed") { + + when { + process { + """ + input[0] = [ + [ + id: 'test_genome', + accession: 'SRR123456', + assembly_software: 'SPAdes_v3.15.5', + binning_software: 'MetaBAT2_v2.15', + binning_parameters: '--minContig 1500', + stats_generation_software: 'CheckM2_v1.0.0', + completeness: '98.4', + contamination: '1.2', + genome_coverage: '42.7', + metagenome: 'marine metagenome', + co_assembly: true, + broad_environment: 'marine biome', + local_environment: 'coastal water', + environmental_medium: 'seawater', + RNA_presence: false, NCBI_lineage: 'Bacteria;Proteobacteria;Gammaproteobacteria' ], file('https://raw.githubusercontent.com/nf-core/test-datasets/viralrecon/genome/NC_045512.2/GCF_009858895.2_ASM985889v3_genomic.200409.fna.gz') @@ -65,11 +104,11 @@ nextflow_process { contamination: '1.2', genome_coverage: '42.7', metagenome: 'marine metagenome', - co_assembly: 'No', + co_assembly: true, broad_environment: 'marine biome', local_environment: 'coastal water', environmental_medium: 'seawater', - RNA_presence: 'Yes', + RNA_presence: false, NCBI_lineage: 'Bacteria;Proteobacteria;Gammaproteobacteria' ], file('https://raw.githubusercontent.com/nf-core/test-datasets/viralrecon/genome/NC_045512.2/GCF_009858895.2_ASM985889v3_genomic.200409.fna.gz') diff --git a/modules/local/create_genome_metadata_tsv/tests/main.nf.test.snap b/modules/local/create_genome_metadata_tsv/tests/main.nf.test.snap index a073053..1de783c 100644 --- a/modules/local/create_genome_metadata_tsv/tests/main.nf.test.snap +++ b/modules/local/create_genome_metadata_tsv/tests/main.nf.test.snap @@ -1,4 +1,68 @@ { + "test_create_genome_metadata_tsv_bool_reversed": { + "content": [ + { + "0": [ + [ + { + "id": "test_genome", + "accession": "SRR123456", + "assembly_software": "SPAdes_v3.15.5", + "binning_software": "MetaBAT2_v2.15", + "binning_parameters": "--minContig 1500", + "stats_generation_software": "CheckM2_v1.0.0", + "completeness": "98.4", + "contamination": "1.2", + "genome_coverage": "42.7", + "metagenome": "marine metagenome", + "co_assembly": true, + "broad_environment": "marine biome", + "local_environment": "coastal water", + "environmental_medium": "seawater", + "RNA_presence": false, + "NCBI_lineage": "Bacteria;Proteobacteria;Gammaproteobacteria" + }, + "test_genome_genome_metadata.tsv:md5,0f955fb3bbf7da09ce9f449884693806" + ] + ], + "1": [ + [ + "CREATE_GENOME_METADATA_TSV", + "bash", + "5.2.37(1)-release" + ] + ], + "tsv": [ + [ + { + "id": "test_genome", + "accession": "SRR123456", + "assembly_software": "SPAdes_v3.15.5", + "binning_software": "MetaBAT2_v2.15", + "binning_parameters": "--minContig 1500", + "stats_generation_software": "CheckM2_v1.0.0", + "completeness": "98.4", + "contamination": "1.2", + "genome_coverage": "42.7", + "metagenome": "marine metagenome", + "co_assembly": true, + "broad_environment": "marine biome", + "local_environment": "coastal water", + "environmental_medium": "seawater", + "RNA_presence": false, + "NCBI_lineage": "Bacteria;Proteobacteria;Gammaproteobacteria" + }, + "test_genome_genome_metadata.tsv:md5,0f955fb3bbf7da09ce9f449884693806" + ] + ] + } + ], + "timestamp": "2026-07-23T14:00:57.58563", + "meta": { + "nf-test": "0.9.5", + "nextflow": "26.04.0" + } + }, "test_create_genome_metadata_tsv - stub": { "content": [ { @@ -15,11 +79,11 @@ "contamination": "1.2", "genome_coverage": "42.7", "metagenome": "marine metagenome", - "co_assembly": "No", + "co_assembly": true, "broad_environment": "marine biome", "local_environment": "coastal water", "environmental_medium": "seawater", - "RNA_presence": "Yes", + "RNA_presence": false, "NCBI_lineage": "Bacteria;Proteobacteria;Gammaproteobacteria" }, "test_genome_genome_metadata.tsv:md5,d41d8cd98f00b204e9800998ecf8427e" @@ -45,11 +109,11 @@ "contamination": "1.2", "genome_coverage": "42.7", "metagenome": "marine metagenome", - "co_assembly": "No", + "co_assembly": true, "broad_environment": "marine biome", "local_environment": "coastal water", "environmental_medium": "seawater", - "RNA_presence": "Yes", + "RNA_presence": false, "NCBI_lineage": "Bacteria;Proteobacteria;Gammaproteobacteria" }, "test_genome_genome_metadata.tsv:md5,d41d8cd98f00b204e9800998ecf8427e" @@ -57,7 +121,7 @@ ] } ], - "timestamp": "2026-07-07T13:49:45.599115", + "timestamp": "2026-07-22T23:11:26.015229", "meta": { "nf-test": "0.9.5", "nextflow": "26.04.0" @@ -79,11 +143,11 @@ "contamination": "1.2", "genome_coverage": "42.7", "metagenome": "marine metagenome", - "co_assembly": "No", + "co_assembly": false, "broad_environment": "marine biome", "local_environment": "coastal water", "environmental_medium": "seawater", - "RNA_presence": "Yes", + "RNA_presence": true, "NCBI_lineage": "Bacteria;Proteobacteria;Gammaproteobacteria" }, "test_genome_genome_metadata.tsv:md5,64562a85d19fe5893cad0e89e894058a" @@ -109,11 +173,11 @@ "contamination": "1.2", "genome_coverage": "42.7", "metagenome": "marine metagenome", - "co_assembly": "No", + "co_assembly": false, "broad_environment": "marine biome", "local_environment": "coastal water", "environmental_medium": "seawater", - "RNA_presence": "Yes", + "RNA_presence": true, "NCBI_lineage": "Bacteria;Proteobacteria;Gammaproteobacteria" }, "test_genome_genome_metadata.tsv:md5,64562a85d19fe5893cad0e89e894058a" @@ -121,7 +185,7 @@ ] } ], - "timestamp": "2026-07-07T13:49:37.674712", + "timestamp": "2026-07-22T23:11:19.939886", "meta": { "nf-test": "0.9.5", "nextflow": "26.04.0" diff --git a/modules/local/generate_assembly_manifest/tests/main.nf.test b/modules/local/generate_assembly_manifest/tests/main.nf.test index 7e9c3bf..e16d278 100644 --- a/modules/local/generate_assembly_manifest/tests/main.nf.test +++ b/modules/local/generate_assembly_manifest/tests/main.nf.test @@ -12,17 +12,16 @@ nextflow_process { when { process { """ - input[0] = [ + input[0] = Channel.of([ [ id:'test' ], file(params.pipelines_testdata_base_path + "/test_data/assembly/contigs.fasta.gz", checkIfExists: true), file(params.pipelines_testdata_base_path + "/samplesheets/samplesheet_generatemanifest.csv", checkIfExists: true) - ] + ]) input[1] = "PRJ12345" input[2] = false input[3] = true input[4] = false """ - } } @@ -30,9 +29,6 @@ nextflow_process { assert process.success assertAll( // Only snapshot versions, not the manifest because it contains a timestamp - { assert snapshot( - path(process.out.versions[0]).yaml - ).match() }, { assert process.out.manifest.size() == 1 }, { assert process.out.manifest[0][1].toString().endsWith(".manifest") }, { @@ -76,8 +72,7 @@ nextflow_process { assert process.success assertAll( { assert snapshot( - process.out, - path(process.out.versions[0]).yaml + process.out ).match() } ) } diff --git a/modules/local/generate_assembly_manifest/tests/main.nf.test.snap b/modules/local/generate_assembly_manifest/tests/main.nf.test.snap index 7d4d1ef..5a6e766 100644 --- a/modules/local/generate_assembly_manifest/tests/main.nf.test.snap +++ b/modules/local/generate_assembly_manifest/tests/main.nf.test.snap @@ -11,7 +11,11 @@ ] ], "1": [ - "versions.yml:md5,5f33d074e4f7f7ddf4b3865973cb9157" + [ + "GENERATE_ASSEMBLY_MANIFEST", + "assembly_uploader", + "1.3.5" + ] ], "manifest": [ [ @@ -20,18 +24,10 @@ }, "test.manifest:md5,d41d8cd98f00b204e9800998ecf8427e" ] - ], - "versions": [ - "versions.yml:md5,5f33d074e4f7f7ddf4b3865973cb9157" ] - }, - { - "GENERATE_ASSEMBLY_MANIFEST": { - "assembly_uploader": "1.3.5" - } } ], - "timestamp": "2026-06-25T22:01:14.155809", + "timestamp": "2026-07-22T21:23:02.99938", "meta": { "nf-test": "0.9.5", "nextflow": "26.04.0" diff --git a/workflows/assemblysubmit.nf b/workflows/assemblysubmit.nf index 401a32f..90273ab 100644 --- a/workflows/assemblysubmit.nf +++ b/workflows/assemblysubmit.nf @@ -51,7 +51,7 @@ workflow ASSEMBLYSUBMIT { // --------- Create assembly channel with proper metadata structure assembly_fasta = ch_samplesheet .map { meta, fasta, reads_1, reads_2 -> - def new_meta = meta + [single_end: reads_2 == null] + def new_meta = meta + [single_end: !reads_2] [new_meta, fasta] } @@ -59,14 +59,11 @@ workflow ASSEMBLYSUBMIT { reads_fastq = ch_samplesheet .filter { row -> row[2] && row[2] != "" } // Check if fastq_1 exists and is not empty .map { meta, fasta, reads_1, reads_2 -> - def new_meta = meta + [single_end: reads_2 == null] - if ( !new_meta.single_end ) { - // If paired end reads - [new_meta, [file(reads_1), file(reads_2)]] - } else { - // If single end - [new_meta, file(reads_1)] - } + def new_meta = meta + [single_end: !reads_2] + def reads = new_meta.single_end + ? [reads_1] + : [reads_1, reads_2] + [new_meta, reads] } // --------- Check fasta files are properly formatted and filter out files with less than 2 contigs diff --git a/workflows/genomesubmit.nf b/workflows/genomesubmit.nf index be4cec9..2cc74b3 100644 --- a/workflows/genomesubmit.nf +++ b/workflows/genomesubmit.nf @@ -62,14 +62,11 @@ workflow GENOMESUBMIT { // --------- Create genomes channel with proper metadata structure genome_fasta_and_reads = ch_samplesheet .map { meta, fasta, reads_1, reads_2 -> - def new_meta = meta + [single_end: reads_2 == null] - if ( !new_meta.single_end ) { - // If paired end reads - return [new_meta, file(fasta), [reads_1, reads_2]] - } else { - // If single end - return [new_meta, file(fasta), [reads_1]] - } + def new_meta = meta + [single_end: !reads_2] + def reads = new_meta.single_end + ? [reads_1] + : [reads_1, reads_2] + [new_meta, fasta, reads] } genome_fasta = genome_fasta_and_reads.map{meta, fasta, _fq1 -> [meta, fasta]} @@ -117,7 +114,7 @@ workflow GENOMESUBMIT { // --------- For genomes without RNA_presence info, calculate rRNA and tRNA branched_rna_results = fasta_updated_with_coverage .branch { meta, _fasta -> - rna_prediction_input: !(meta.RNA_presence) + rna_prediction_input: meta.RNA_presence == null // it might be True/False rna_present: true // Everything else goes here } @@ -132,7 +129,7 @@ workflow GENOMESUBMIT { .map{ meta, rna_decision, fasta -> def lines = rna_decision.readLines() // support for empty decision files required for -stub mode - def decision = lines ? lines[0].split('\t')[1] : null + def decision = lines ? lines[0].split('\t')[1].toLowerCase() == 'true' : null def updated_meta = meta.clone() updated_meta.RNA_presence = decision return [updated_meta, fasta] diff --git a/workflows/readsubmit.nf b/workflows/readsubmit.nf index 45c375e..8bdff88 100644 --- a/workflows/readsubmit.nf +++ b/workflows/readsubmit.nf @@ -46,13 +46,13 @@ workflow READSUBMIT { // --------- Create reads channel with proper metadata structure reads_ch = ch_samplesheet .map { meta, reads_1, reads_2 -> - def new_meta = meta + [single_end: reads_2 == null] + def new_meta = meta + [single_end: !reads_2] if ( !new_meta.single_end ) { // If paired end reads - [new_meta, [file(reads_1), file(reads_2)]] + [new_meta, [reads_1, reads_2]] } else { // If single end - [new_meta, file(reads_1)] + [new_meta, reads_1] } } From d3d61ad663fd36b251c841b5b745b38439d1ffd5 Mon Sep 17 00:00:00 2001 From: nf-core-bot Date: Fri, 24 Jul 2026 12:36:31 +0000 Subject: [PATCH 36/47] Template update for nf-core/tools version 4.0.3 --- .github/PULL_REQUEST_TEMPLATE.md | 4 +- .github/actions/nf-test/action.yml | 2 +- .github/workflows/branch.yml | 60 +++++++++----- .github/workflows/linting.yml | 18 ++++ .github/workflows/linting_comment.yml | 28 ------- .github/workflows/nf-test.yml | 57 +++++++++++++ .github/workflows/pr-comment.yml | 82 +++++++++++++++++++ .github/workflows/release-announcements.yml | 2 +- .../workflows/template-version-comment.yml | 56 ++++++++----- .nf-core.yml | 8 +- CHANGELOG.md | 2 +- README.md | 4 +- assets/multiqc_config.yml | 4 +- assets/schema_input.json | 2 +- nextflow.config | 14 +++- nextflow_schema.json | 2 +- ro-crate-metadata.json | 38 +++++---- .../utils_nfcore_seqsubmit_pipeline/main.nf | 2 +- 18 files changed, 281 insertions(+), 104 deletions(-) delete mode 100644 .github/workflows/linting_comment.yml create mode 100644 .github/workflows/pr-comment.yml diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md index c8ad298..d3e7af9 100644 --- a/.github/PULL_REQUEST_TEMPLATE.md +++ b/.github/PULL_REQUEST_TEMPLATE.md @@ -8,14 +8,14 @@ These are the most common things requested on pull requests (PRs). Remember that PRs should be made against the dev branch, unless you're preparing a pipeline release. -Learn more about contributing: [CONTRIBUTING.md](https://github.com/nf-core/seqsubmit/tree/master/docs/CONTRIBUTING.md) +Learn more about contributing: [CONTRIBUTING.md](https://github.com/nf-core/seqsubmit/tree/main/docs/CONTRIBUTING.md) --> ## PR checklist - [ ] This comment contains a description of changes (with reason). - [ ] If you've fixed a bug or added code that should be tested, add tests! -- [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/seqsubmit/tree/master/docs/CONTRIBUTING.md) +- [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/seqsubmit/tree/main/docs/CONTRIBUTING.md) - [ ] If necessary, also make a PR on the nf-core/seqsubmit _branch_ on the [nf-core/test-datasets](https://github.com/nf-core/test-datasets) repository. - [ ] Make sure your code lints (`nf-core pipelines lint`). - [ ] Ensure the test suite passes (`nextflow run . -profile test,docker --outdir `). diff --git a/.github/actions/nf-test/action.yml b/.github/actions/nf-test/action.yml index ad686e8..945c56f 100644 --- a/.github/actions/nf-test/action.yml +++ b/.github/actions/nf-test/action.yml @@ -37,7 +37,7 @@ runs: - name: Setup apptainer if: contains(inputs.profile, 'singularity') - uses: eWaterCycle/setup-apptainer@3f706d898c9db585b1d741b4692e66755f3a1b40 # v2 + uses: eWaterCycle/setup-apptainer@4bb22c52d4f63406c49e94c804632975787312b3 # v2.0.0 - name: Set up Singularity if: contains(inputs.profile, 'singularity') diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml index 7bbd3f2..9075dd6 100644 --- a/.github/workflows/branch.yml +++ b/.github/workflows/branch.yml @@ -2,11 +2,13 @@ name: nf-core branch protection # This workflow is triggered on PRs to `main`/`master` branch on the repository # It fails when someone tries to make a PR against the nf-core `main`/`master` branch instead of `dev` on: - pull_request_target: + pull_request: branches: - main - master +permissions: {} + jobs: test: runs-on: ubuntu-latest @@ -14,33 +16,47 @@ jobs: # PRs to the nf-core repo main/master branch are only ok if coming from the nf-core repo `dev` or any `patch` branches - name: Check PRs if: github.repository == 'nf-core/seqsubmit' + env: + HEAD_REPO: ${{ github.event.pull_request.head.repo.full_name }} run: | - { [[ ${{github.event.pull_request.head.repo.full_name }} == nf-core/seqsubmit ]] && [[ $GITHUB_HEAD_REF == "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] + { [[ "$HEAD_REPO" == nf-core/seqsubmit ]] && [[ $GITHUB_HEAD_REF == "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] - # If the above check failed, post a comment on the PR explaining the failure - # NOTE - this doesn't currently work if the PR is coming from a fork, due to limitations in GitHub actions secrets - - name: Post PR comment + # If the above check failed, build a comment to be posted by the shared poster workflow + - name: Build PR comment if: failure() - uses: mshick/add-pr-comment@8e4927817251f1ff60c001f04568532b38e0b4a0 # v3 - with: - message: | - ## This PR is against the `${{github.event.pull_request.base.ref}}` branch :x: + env: + PR_NUMBER: ${{ github.event.pull_request.number }} + BASE_REF: ${{ github.event.pull_request.base.ref }} + HEAD_REPO: ${{ github.event.pull_request.head.repo.full_name }} + PR_USER: ${{ github.event.pull_request.user.login }} + run: | + mkdir -p pr-comment + echo "$PR_NUMBER" > pr-comment/pr_number.txt + echo "branch" > pr-comment/header.txt + cat > pr-comment/comment.md < pr-comment/pr_number.txt + echo "lint" > pr-comment/header.txt + [ -f lint_results.md ] && cp lint_results.md pr-comment/comment.md || true + + - name: Upload PR comment artifact + if: ${{ always() }} + uses: actions/upload-artifact@043fb46d1a93c77aae656e7c1c64a875d1fc6a0a # v7 + with: + name: pr-comment + path: pr-comment/ diff --git a/.github/workflows/linting_comment.yml b/.github/workflows/linting_comment.yml deleted file mode 100644 index 5b0c24f..0000000 --- a/.github/workflows/linting_comment.yml +++ /dev/null @@ -1,28 +0,0 @@ -name: nf-core linting comment -# This workflow is triggered after the linting action is complete -# It posts an automated comment to the PR, even if the PR is coming from a fork - -on: - workflow_run: - workflows: ["nf-core linting"] - -jobs: - test: - runs-on: ubuntu-latest - steps: - - name: Download lint results - uses: dawidd6/action-download-artifact@b6e2e70617bc3265edd6dab6c906732b2f1ae151 # v21 - with: - workflow: linting.yml - workflow_conclusion: completed - - - name: Get PR number - id: pr_number - run: echo "pr_number=$(cat linting-logs/PR_number.txt)" >> $GITHUB_OUTPUT - - - name: Post PR comment - uses: marocchino/sticky-pull-request-comment@70d2764d1a7d5d9560b100cbea0077fc8f633987 # v3 - with: - GITHUB_TOKEN: ${{ secrets.GITHUB_TOKEN }} - number: ${{ steps.pr_number.outputs.pr_number }} - path: linting-logs/lint_results.md diff --git a/.github/workflows/nf-test.yml b/.github/workflows/nf-test.yml index efd72d6..4de681a 100644 --- a/.github/workflows/nf-test.yml +++ b/.github/workflows/nf-test.yml @@ -116,6 +116,22 @@ jobs: fi fi + # continue-on-error keeps latest-everything from failing the job, so it never shows up in + # `needs.nf-test.result` downstream and CI stays green. Surface it via a PR comment instead; + # other NXF_VER failures already fail the job/CI directly, so no comment is needed for those. + - name: Prepare PR comment fragment + if: ${{ always() && steps.run_nf_test.outcome == 'failure' && matrix.NXF_VER == 'latest-everything' }} + run: | + mkdir -p pr-comment-fragment + echo "* ❌ \`${{ matrix.profile }}\` | \`${{ matrix.NXF_VER }}\` | Shard ${{ matrix.shard }}/${{ env.TOTAL_SHARDS }}" > pr-comment-fragment/fragment.md + + - name: Upload PR comment fragment + if: ${{ always() && steps.run_nf_test.outcome == 'failure' && matrix.NXF_VER == 'latest-everything' }} + uses: actions/upload-artifact@043fb46d1a93c77aae656e7c1c64a875d1fc6a0a # v7 + with: + name: pr-comment-fragment-${{ strategy.job-index }} + path: pr-comment-fragment/ + confirm-pass: needs: [nf-test] if: always() @@ -142,3 +158,44 @@ jobs: echo "DEBUG: toJSON(needs) = ${{ toJSON(needs) }}" echo "DEBUG: toJSON(needs.*.result) = ${{ toJSON(needs.*.result) }}" echo "::endgroup::" + + - name: Download PR comment fragments + if: ${{ always() }} + uses: actions/download-artifact@3e5f45b2cfb9172054b4087a40e8e0b5a5461e7c # v8.0.1 + continue-on-error: true + with: + pattern: pr-comment-fragment-* + path: pr-comment-fragments + merge-multiple: true + + # Build a comment for the shared pr-comment.yml poster to publish on the PR. + # Based on the fragments above (not needs.*.result) so non-blocking failures are still reported. + - name: Prepare PR comment + if: ${{ always() }} + env: + PR_NUMBER: ${{ github.event.pull_request.number }} + RUN_URL: ${{ github.server_url }}/${{ github.repository }}/actions/runs/${{ github.run_id }} + run: | + mkdir -p pr-comment + echo "$PR_NUMBER" > pr-comment/pr_number.txt + echo "nf-test" > pr-comment/header.txt + if [ -d pr-comment-fragments ] && [ -n "$(ls -A pr-comment-fragments)" ]; then + { + echo "## ❌ nf-test failed with latest Nextflow version" + echo "" + echo "> [!NOTE]" + echo "> Tests with Nextflow's latest version failed but it will not cause a CI workflow failure." + echo "> Please check if the failure is expected with newer (edge-)releases of Nextflow or if it needs fixing." + echo "" + cat pr-comment-fragments/*.md + echo "" + echo "See the [full run](${RUN_URL}) for details." + } > pr-comment/comment.md + fi + + - name: Upload PR comment artifact + if: ${{ always() }} + uses: actions/upload-artifact@043fb46d1a93c77aae656e7c1c64a875d1fc6a0a # v7 + with: + name: pr-comment + path: pr-comment/ diff --git a/.github/workflows/pr-comment.yml b/.github/workflows/pr-comment.yml new file mode 100644 index 0000000..ab7b59d --- /dev/null +++ b/.github/workflows/pr-comment.yml @@ -0,0 +1,82 @@ +name: Post PR comment +# Shared, privileged comment poster. +# +# This is the single workflow that runs with a write token. It is triggered +# after any of the listed "producer" workflows complete on a pull request. +# Each producer runs untrusted PR code (if any) with a read-only token and +# uploads a `pr-comment` artifact describing the comment to post; this workflow +# only ever reads that plain-text artifact, so no PR code is executed here. +# +# Artifact contract (uploaded by producers under the name `pr-comment`): +# pr_number.txt - the pull request number +# header.txt - sticky-comment identifier (keeps comment types separate) +# comment.md - the Markdown body (omit the file to post nothing) + +on: + workflow_run: + workflows: + - "nf-core linting" + - "nf-core template version comment" + - "nf-core branch protection" + - "Run nf-test" + +permissions: + actions: read + contents: read + pull-requests: write + +jobs: + post-comment: + runs-on: ubuntu-latest + if: github.event.workflow_run.event == 'pull_request' + steps: + - name: Download PR comment artifact + uses: dawidd6/action-download-artifact@b6e2e70617bc3265edd6dab6c906732b2f1ae151 # v21 + with: + run_id: ${{ github.event.workflow_run.id }} + name: pr-comment + path: pr-comment + if_no_artifact_found: ignore + + - name: Read comment metadata + id: meta + run: | + echo "::group::Downloaded pr-comment contents" + ls -la pr-comment 2>/dev/null || echo "No pr-comment/ directory was downloaded." + echo "::endgroup::" + + if [ ! -d pr-comment ]; then + echo "No pr-comment artifact found; nothing to post." + exit 0 + fi + + if [ ! -f pr-comment/comment.md ]; then + echo "Artifact present but no comment.md; nothing to post." + exit 0 + fi + + pr_number=$(cat pr-comment/pr_number.txt) + header=$(cat pr-comment/header.txt) + echo "Found comment.md (header='$header', pr_number='$pr_number')." + + # Guard against anything unexpected ending up in the PR number. + case "$pr_number" in + ''|*[!0-9]*) + echo "Invalid PR number: '$pr_number'" + exit 1 + ;; + esac + + echo "pr_number=$pr_number" >> "$GITHUB_OUTPUT" + echo "header=$header" >> "$GITHUB_OUTPUT" + echo "post=true" >> "$GITHUB_OUTPUT" + echo "Will post comment to PR #${pr_number}." + + - name: Post PR comment + if: steps.meta.outputs.post == 'true' + uses: marocchino/sticky-pull-request-comment@5770ad5eb8f42dd2c4f34da00c94c5381e49af88 # v3.0.5 + with: + GITHUB_TOKEN: ${{ secrets.GITHUB_TOKEN }} + number: ${{ steps.meta.outputs.pr_number }} + header: ${{ steps.meta.outputs.header }} + path: pr-comment/comment.md diff --git a/.github/workflows/release-announcements.yml b/.github/workflows/release-announcements.yml index 78d5dbe..4974f44 100644 --- a/.github/workflows/release-announcements.yml +++ b/.github/workflows/release-announcements.yml @@ -18,7 +18,7 @@ jobs: id: get_description run: | echo "description=$(curl -s https://nf-co.re/pipelines.json | jq -r '.remote_workflows[] | select(.full_name == "${{ github.repository }}") | .description')" >> $GITHUB_OUTPUT - - uses: rzr/fediverse-action@563159eb8d45f70ab6aaba36ed55cd037e51f441 # master + - uses: rzr/fediverse-action@66c2cbb5b1997666b0e28d597631b6a4f09a2719 # v0.0.6 with: access-token: ${{ secrets.MASTODON_ACCESS_TOKEN }} host: "mstdn.science" # custom host if not "mastodon.social" (default) diff --git a/.github/workflows/template-version-comment.yml b/.github/workflows/template-version-comment.yml index ea30827..ee102f7 100644 --- a/.github/workflows/template-version-comment.yml +++ b/.github/workflows/template-version-comment.yml @@ -2,10 +2,13 @@ name: nf-core template version comment # This workflow is triggered on PRs to check if the pipeline template version matches the latest nf-core version. # It posts a comment to the PR, even if it comes from a fork. -on: pull_request_target +on: + pull_request: + +permissions: {} jobs: - template_version: + check_template_version: runs-on: ubuntu-latest steps: - name: Check out pipeline code @@ -22,25 +25,36 @@ jobs: - name: Install nf-core run: | python -m pip install --upgrade pip - pip install nf-core==${{ steps.read_yml.outputs['nf_core_version'] }} + pip install nf-core + + - name: Build PR comment if template is outdated + # The fork-controlled version is passed via the environment and only ever + # used as quoted shell data (never interpolated into a command), so it + # cannot be used for script injection. + env: + PR_VERSION: ${{ steps.read_yml.outputs['nf_core_version'] }} + PR_NUMBER: ${{ github.event.pull_request.number }} + run: | + mkdir -p pr-comment + echo "$PR_NUMBER" > pr-comment/pr_number.txt + echo "template-version" > pr-comment/header.txt + + latest_version=$(nf-core --version | grep -oE '[0-9]+\.[0-9]+\.[0-9]+' | head -n1) - - name: Check nf-core outdated - id: nf_core_outdated - run: echo "OUTPUT=$(pip list --outdated | grep nf-core)" >> ${GITHUB_ENV} + if [ -n "$PR_VERSION" ] && [ -n "$latest_version" ] && [ "$PR_VERSION" != "$latest_version" ]; then + cat > pr-comment/comment.md < [!WARNING] + > Newer version of the nf-core template is available. + > + > Your pipeline is using an old version of the nf-core template: ${PR_VERSION}. + > Please update your pipeline to the latest version. + > + > For more documentation on how to update your pipeline, please see the [Synchronisation documentation](https://nf-co.re/docs/developing/template-syncs/overview). + EOF + fi - - name: Post nf-core template version comment - uses: mshick/add-pr-comment@8e4927817251f1ff60c001f04568532b38e0b4a0 # v3 - if: | - contains(env.OUTPUT, 'nf-core') + - name: Upload PR comment artifact + uses: actions/upload-artifact@043fb46d1a93c77aae656e7c1c64a875d1fc6a0a # v7 with: - repo-token: ${{ secrets.NF_CORE_BOT_AUTH_TOKEN }} - allow-repeats: false - message: | - > [!WARNING] - > Newer version of the nf-core template is available. - > - > Your pipeline is using an old version of the nf-core template: ${{ steps.read_yml.outputs['nf_core_version'] }}. - > Please update your pipeline to the latest version. - > - > For more documentation on how to update your pipeline, please see the [Synchronisation documentation](https://nf-co.re/docs/developing/template-syncs/overview). - # + name: pr-comment + path: pr-comment/ diff --git a/.nf-core.yml b/.nf-core.yml index fcd768a..9eff1ef 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -1,4 +1,5 @@ lint: + container_configs: false files_exist: - conf/igenomes.config - conf/igenomes_ignored.config @@ -9,10 +10,11 @@ lint: - assets/nf-core-seqsubmit_logo_light.png nextflow_config: - params.input -nf_core_version: 4.0.2 +nf_core_version: 4.0.3 repository_type: pipeline template: - author: Martin Beracochea, Ekaterina Sakharova, Sofiia Ochkalova, Evangelos Karatzas + author: Martin Beracochea, Ekaterina Sakharova, Sofia Ochkalova, Evangelos Karatzas, + Tim Rozday description: Submit sequences, assemblies, MAGs, etc., to public archives force: false is_nfcore: true @@ -23,4 +25,4 @@ template: - igenomes - fastqc - gpu - version: 1.0.0dev + version: 1.0.0 diff --git a/CHANGELOG.md b/CHANGELOG.md index 8859bd8..90900cd 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). -## v1.0.0dev - [date] +## v1.0.0 - [date] Initial release of nf-core/seqsubmit, created with the [nf-core](https://nf-co.re/) template. diff --git a/README.md b/README.md index 0e9106d..59ccc3c 100644 --- a/README.md +++ b/README.md @@ -11,7 +11,7 @@ [![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com) [![Nextflow](https://img.shields.io/badge/version-%E2%89%A525.10.4-green?style=flat&logo=nextflow&logoColor=white&color=%230DC09D&link=https%3A%2F%2Fnextflow.io)](https://www.nextflow.io/) -[![nf-core template version](https://img.shields.io/badge/nf--core_template-4.0.2-green?style=flat&logo=nfcore&logoColor=white&color=%2324B064&link=https%3A%2F%2Fnf-co.re)](https://github.com/nf-core/tools/releases/tag/4.0.2) +[![nf-core template version](https://img.shields.io/badge/nf--core_template-4.0.3-green?style=flat&logo=nfcore&logoColor=white&color=%2324B064&link=https%3A%2F%2Fnf-co.re)](https://github.com/nf-core/tools/releases/tag/4.0.3) [![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/) [![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/) [![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/) @@ -78,7 +78,7 @@ For more details about the output files and reports, please refer to the ## Credits -nf-core/seqsubmit was originally written by Martin Beracochea, Ekaterina Sakharova, Sofiia Ochkalova, Evangelos Karatzas. +nf-core/seqsubmit was originally written by Martin Beracochea, Ekaterina Sakharova, Sofia Ochkalova, Evangelos Karatzas, Tim Rozday. We thank the following people for their extensive assistance in the development of this pipeline: diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index 5b6dfbc..66a4f8c 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -1,7 +1,7 @@ report_comment: > - This report has been generated by the     nf-core/seqsubmit + This report has been generated by the nf-core/seqsubmit analysis pipeline. For information about how to interpret these results, please see the - documentation. + documentation. report_section_order: "nf-core-seqsubmit-methods-description": order: -1000 diff --git a/assets/schema_input.json b/assets/schema_input.json index e5c86ba..4d8c527 100644 --- a/assets/schema_input.json +++ b/assets/schema_input.json @@ -1,6 +1,6 @@ { "$schema": "https://json-schema.org/draft/2020-12/schema", - "$id": "https://raw.githubusercontent.com/nf-core/seqsubmit/master/assets/schema_input.json", + "$id": "https://raw.githubusercontent.com/nf-core/seqsubmit/main/assets/schema_input.json", "title": "nf-core/seqsubmit pipeline - params.input schema", "description": "Schema for the file provided with params.input", "type": "array", diff --git a/nextflow.config b/nextflow.config index adc4028..8ec9406 100644 --- a/nextflow.config +++ b/nextflow.config @@ -252,7 +252,7 @@ manifest { orcid: '' ], [ - name: ' Sofiia Ochkalova', + name: ' Sofia Ochkalova', affiliation: '', email: '', github: '', @@ -267,13 +267,21 @@ manifest { contribution: [], // List of contribution types ('author', 'maintainer' or 'contributor') orcid: '' ], + [ + name: ' Tim Rozday', + affiliation: '', + email: '', + github: '', + contribution: [], // List of contribution types ('author', 'maintainer' or 'contributor') + orcid: '' + ], ] homePage = 'https://github.com/nf-core/seqsubmit' description = """Submit sequences, assemblies, MAGs, etc., to public archives""" mainScript = 'main.nf' - defaultBranch = 'master' + defaultBranch = 'main' nextflowVersion = '!>=25.10.4' - version = '1.0.0dev' + version = '1.0.0' doi = '' } diff --git a/nextflow_schema.json b/nextflow_schema.json index 7a4b039..9785121 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -1,6 +1,6 @@ { "$schema": "https://json-schema.org/draft/2020-12/schema", - "$id": "https://raw.githubusercontent.com/nf-core/seqsubmit/master/nextflow_schema.json", + "$id": "https://raw.githubusercontent.com/nf-core/seqsubmit/main/nextflow_schema.json", "title": "nf-core/seqsubmit pipeline parameters", "description": "Submit sequences, assemblies, MAGs, etc., to public archives", "type": "object", diff --git a/ro-crate-metadata.json b/ro-crate-metadata.json index ca98c1b..c13fe1d 100644 --- a/ro-crate-metadata.json +++ b/ro-crate-metadata.json @@ -21,9 +21,9 @@ { "@id": "./", "@type": "Dataset", - "creativeWorkStatus": "InProgress", - "datePublished": "2026-04-30T13:32:49+00:00", - "description": "

    \n \n \n \"nf-core/seqsubmit\"\n \n

    \n\n[![Open in GitHub Codespaces](https://img.shields.io/badge/Open_In_GitHub_Codespaces-black?labelColor=grey&logo=github)](https://github.com/codespaces/new/nf-core/seqsubmit)\n[![GitHub Actions CI Status](https://github.com/nf-core/seqsubmit/actions/workflows/nf-test.yml/badge.svg)](https://github.com/nf-core/seqsubmit/actions/workflows/nf-test.yml)\n[![GitHub Actions Linting Status](https://github.com/nf-core/seqsubmit/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/seqsubmit/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/seqsubmit/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX)\n[![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com)\n\n[![Nextflow](https://img.shields.io/badge/version-%E2%89%A525.10.4-green?style=flat&logo=nextflow&logoColor=white&color=%230DC09D&link=https%3A%2F%2Fnextflow.io)](https://www.nextflow.io/)\n[![nf-core template version](https://img.shields.io/badge/nf--core_template-4.0.2-green?style=flat&logo=nfcore&logoColor=white&color=%2324B064&link=https%3A%2F%2Fnf-co.re)](https://github.com/nf-core/tools/releases/tag/4.0.2)\n[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/)\n[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/)\n[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/)\n[![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/seqsubmit)\n\n[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23seqsubmit-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/seqsubmit)[![Follow on Bluesky](https://img.shields.io/badge/bluesky-%40nf__core-1185fe?labelColor=000000&logo=bluesky)](https://bsky.app/profile/nf-co.re)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core)\n\n## Introduction\n\n**nf-core/seqsubmit** is a bioinformatics pipeline that ...\n\n\n\n\n2. Present QC for raw reads ([`MultiQC`](http://multiqc.info/))\n\n## Usage\n\n> [!NOTE]\n> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/get_started/environment_setup/overview) on how to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/get_started/run-your-first-pipeline) with `-profile test` before running the workflow on actual data.\n\n\n\nNow, you can run the pipeline using:\n\n\n\n```bash\nnextflow run nf-core/seqsubmit \\\n -profile \\\n --input samplesheet.csv \\\n --outdir \n```\n\n> [!WARNING]\n> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; see [docs](https://nf-co.re/docs/running/run-pipelines#using-parameter-files).\n\nFor more details and further functionality, please refer to the [usage documentation](https://nf-co.re/seqsubmit/usage) and the [parameter documentation](https://nf-co.re/seqsubmit/parameters).\n\n## Pipeline output\n\nTo see the results of an example test run with a full size dataset refer to the [results](https://nf-co.re/seqsubmit/results) tab on the nf-core website pipeline page.\nFor more details about the output files and reports, please refer to the\n[output documentation](https://nf-co.re/seqsubmit/output).\n\n## Credits\n\nnf-core/seqsubmit was originally written by Martin Beracochea, Ekaterina Sakharova, Sofiia Ochkalova, Evangelos Karatzas.\n\nWe thank the following people for their extensive assistance in the development of this pipeline:\n\n\n\n## Contributions and Support\n\nIf you would like to contribute to this pipeline, please see the [contributing guidelines](docs/CONTRIBUTING.md).\n\nFor further information or help, don't hesitate to get in touch on the [Slack `#seqsubmit` channel](https://nfcore.slack.com/channels/seqsubmit) (you can join with [this invite](https://nf-co.re/join/slack)).\n\n## Citations\n\n\n\n\n\n\nAn extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file.\n\nYou can cite the `nf-core` publication as follows:\n\n> **The nf-core framework for community-curated bioinformatics pipelines.**\n>\n> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen.\n>\n> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x).\n", + "creativeWorkStatus": "Stable", + "datePublished": "2026-07-24T12:36:22+00:00", + "description": "

    \n \n \n \"nf-core/seqsubmit\"\n \n

    \n\n[![Open in GitHub Codespaces](https://img.shields.io/badge/Open_In_GitHub_Codespaces-black?labelColor=grey&logo=github)](https://github.com/codespaces/new/nf-core/seqsubmit)\n[![GitHub Actions CI Status](https://github.com/nf-core/seqsubmit/actions/workflows/nf-test.yml/badge.svg)](https://github.com/nf-core/seqsubmit/actions/workflows/nf-test.yml)\n[![GitHub Actions Linting Status](https://github.com/nf-core/seqsubmit/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/seqsubmit/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/seqsubmit/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX)\n[![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com)\n\n[![Nextflow](https://img.shields.io/badge/version-%E2%89%A525.10.4-green?style=flat&logo=nextflow&logoColor=white&color=%230DC09D&link=https%3A%2F%2Fnextflow.io)](https://www.nextflow.io/)\n[![nf-core template version](https://img.shields.io/badge/nf--core_template-4.0.3-green?style=flat&logo=nfcore&logoColor=white&color=%2324B064&link=https%3A%2F%2Fnf-co.re)](https://github.com/nf-core/tools/releases/tag/4.0.3)\n[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/)\n[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/)\n[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/)\n[![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/seqsubmit)\n\n[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23seqsubmit-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/seqsubmit)[![Follow on Bluesky](https://img.shields.io/badge/bluesky-%40nf__core-1185fe?labelColor=000000&logo=bluesky)](https://bsky.app/profile/nf-co.re)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core)\n\n## Introduction\n\n**nf-core/seqsubmit** is a bioinformatics pipeline that ...\n\n\n\n\n2. Present QC for raw reads ([`MultiQC`](http://multiqc.info/))\n\n## Usage\n\n> [!NOTE]\n> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/get_started/environment_setup/overview) on how to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/get_started/run-your-first-pipeline) with `-profile test` before running the workflow on actual data.\n\n\n\nNow, you can run the pipeline using:\n\n\n\n```bash\nnextflow run nf-core/seqsubmit \\\n -profile \\\n --input samplesheet.csv \\\n --outdir \n```\n\n> [!WARNING]\n> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; see [docs](https://nf-co.re/docs/running/run-pipelines#using-parameter-files).\n\nFor more details and further functionality, please refer to the [usage documentation](https://nf-co.re/seqsubmit/usage) and the [parameter documentation](https://nf-co.re/seqsubmit/parameters).\n\n## Pipeline output\n\nTo see the results of an example test run with a full size dataset refer to the [results](https://nf-co.re/seqsubmit/results) tab on the nf-core website pipeline page.\nFor more details about the output files and reports, please refer to the\n[output documentation](https://nf-co.re/seqsubmit/output).\n\n## Credits\n\nnf-core/seqsubmit was originally written by Martin Beracochea, Ekaterina Sakharova, Sofia Ochkalova, Evangelos Karatzas, Tim Rozday.\n\nWe thank the following people for their extensive assistance in the development of this pipeline:\n\n\n\n## Contributions and Support\n\nIf you would like to contribute to this pipeline, please see the [contributing guidelines](docs/CONTRIBUTING.md).\n\nFor further information or help, don't hesitate to get in touch on the [Slack `#seqsubmit` channel](https://nfcore.slack.com/channels/seqsubmit) (you can join with [this invite](https://nf-co.re/join/slack)).\n\n## Citations\n\n\n\n\n\n\nAn extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file.\n\nYou can cite the `nf-core` publication as follows:\n\n> **The nf-core framework for community-curated bioinformatics pipelines.**\n>\n> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen.\n>\n> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x).\n", "hasPart": [ { "@id": "main.nf" @@ -99,7 +99,7 @@ }, "mentions": [ { - "@id": "#6a35cf24-1f54-4f1b-8bd7-da24e4ef9c61" + "@id": "#b9a22430-cd92-4053-a257-9fd6f20776f9" } ], "name": "nf-core/seqsubmit" @@ -127,17 +127,20 @@ "@id": "https://orcid.org/0000-0003-3472-3736" }, { - "@id": "#002b989f-efa9-4c5b-bf4e-bbf8f2d58be1" + "@id": "#e5eaba7e-b8d4-40dd-a235-ea4a18314307" }, { - "@id": "#bf7a74fc-7f56-4751-bd51-0f3035468776" + "@id": "https://orcid.org/0000-0003-2814-6012" }, { "@id": "https://orcid.org/0000-0001-9132-8981" + }, + { + "@id": "#14ef52aa-83db-4988-82ab-828335934408" } ], "dateCreated": "", - "dateModified": "2026-04-30T13:32:49Z", + "dateModified": "2026-07-24T12:36:22Z", "dct:conformsTo": "https://bioschemas.org/profiles/ComputationalWorkflow/1.0-RELEASE/", "keywords": ["nf-core", "nextflow"], "license": ["MIT"], @@ -148,8 +151,8 @@ "sdPublisher": { "@id": "https://nf-co.re/" }, - "url": ["https://github.com/nf-core/seqsubmit", "https://nf-co.re/seqsubmit/dev/"], - "version": ["1.0.0dev"] + "url": ["https://github.com/nf-core/seqsubmit", "https://nf-co.re/seqsubmit/1.0.0/"], + "version": ["1.0.0"] }, { "@id": "https://w3id.org/workflowhub/workflow-ro-crate#nextflow", @@ -164,11 +167,11 @@ "version": "!>=25.10.4" }, { - "@id": "#6a35cf24-1f54-4f1b-8bd7-da24e4ef9c61", + "@id": "#b9a22430-cd92-4053-a257-9fd6f20776f9", "@type": "TestSuite", "instance": [ { - "@id": "#32731c67-24d2-422c-bacc-c9bf3b9753c3" + "@id": "#4b475711-84b9-4a69-9fa5-544aba88ac60" } ], "mainEntity": { @@ -177,7 +180,7 @@ "name": "Test suite for nf-core/seqsubmit" }, { - "@id": "#32731c67-24d2-422c-bacc-c9bf3b9753c3", + "@id": "#4b475711-84b9-4a69-9fa5-544aba88ac60", "@type": "TestInstance", "name": "GitHub Actions workflow for testing nf-core/seqsubmit", "resource": "repos/nf-core/seqsubmit/actions/workflows/nf-test.yml", @@ -311,19 +314,24 @@ "name": "Martin Beracochea" }, { - "@id": "#002b989f-efa9-4c5b-bf4e-bbf8f2d58be1", + "@id": "#e5eaba7e-b8d4-40dd-a235-ea4a18314307", "@type": "Person", "name": "Ekaterina Sakharova" }, { - "@id": "#bf7a74fc-7f56-4751-bd51-0f3035468776", + "@id": "https://orcid.org/0000-0003-2814-6012", "@type": "Person", - "name": "Sofiia Ochkalova" + "name": "Sofia Ochkalova" }, { "@id": "https://orcid.org/0000-0001-9132-8981", "@type": "Person", "name": "Evangelos Karatzas" + }, + { + "@id": "#14ef52aa-83db-4988-82ab-828335934408", + "@type": "Person", + "name": "Tim Rozday" } ] } diff --git a/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf b/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf index e5a8094..9a0553c 100644 --- a/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_seqsubmit_pipeline/main.nf @@ -71,7 +71,7 @@ workflow PIPELINE_INITIALISATION { https://doi.org/10.1038/s41587-020-0439-x * Software dependencies - https://github.com/nf-core/seqsubmit/blob/master/CITATIONS.md + https://github.com/nf-core/seqsubmit/blob/main/CITATIONS.md """ if (monochrome_logs) { before_text = before_text.replaceAll(/\033\[[0-9;]*m/, '') From d0e42bef1af650ac54ccae6d23f7e1c9e6293058 Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Mon, 27 Jul 2026 11:56:00 +0100 Subject: [PATCH 37/47] change limits for coverage, completeness and contamination --- assets/schema_input_assembly.json | 4 ++-- assets/schema_input_genome.json | 13 +++++++------ 2 files changed, 9 insertions(+), 8 deletions(-) diff --git a/assets/schema_input_assembly.json b/assets/schema_input_assembly.json index c35a887..4a767ca 100644 --- a/assets/schema_input_assembly.json +++ b/assets/schema_input_assembly.json @@ -59,7 +59,7 @@ "anyOf": [ { "type": "number", - "minimum": 0 + "exclusiveMinimum": 0 }, { "type": "string", @@ -108,7 +108,7 @@ "properties": { "coverage": { "type": "number", - "minimum": 0 + "exclusiveMinimum": 0 } }, "required": ["coverage"] diff --git a/assets/schema_input_genome.json b/assets/schema_input_genome.json index 393211d..28c4d69 100644 --- a/assets/schema_input_genome.json +++ b/assets/schema_input_genome.json @@ -40,6 +40,7 @@ "maxLength": 0 } ], + "default": null, "errorMessage": "FASTQ file must have extension '.fq' or '.fastq' (optionally gzipped)", "description": "Forward reads if paired-end or single-end reads FASTQ file" }, @@ -85,8 +86,8 @@ "anyOf": [ { "type": "number", - "exclusiveMinimum": 0, - "exclusiveMaximum": 100 + "minimum": 0, + "maximum": 100 }, { "type": "string", @@ -101,8 +102,8 @@ "anyOf": [ { "type": "number", - "exclusiveMinimum": 0, - "exclusiveMaximum": 100 + "minimum": 0, + "maximum": 100 }, { "type": "string", @@ -117,7 +118,7 @@ "anyOf": [ { "type": "number", - "minimum": 0 + "exclusiveMinimum": 0 }, { "type": "string", @@ -201,7 +202,7 @@ "properties": { "genome_coverage": { "type": "number", - "minimum": 0 + "exclusiveMinimum": 0 } }, "required": ["genome_coverage"] From 774f69c954256cb88444f634def12f06937f6aa7 Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova <49755902+KateSakharova@users.noreply.github.com> Date: Mon, 27 Jul 2026 15:07:47 +0100 Subject: [PATCH 38/47] Apply suggestion from @ochkalova Co-authored-by: Sofia Ochkalova <61541445+ochkalova@users.noreply.github.com> --- conf/test_mag_complete_metadata.conf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test_mag_complete_metadata.conf b/conf/test_mag_complete_metadata.conf index e2d2489..428878a 100644 --- a/conf/test_mag_complete_metadata.conf +++ b/conf/test_mag_complete_metadata.conf @@ -20,7 +20,7 @@ process { params { config_profile_name = 'Test --mode mags complete_metadata profile' - config_profile_description = 'Single-case MAG test with complete metadata in samplesheet. No statistics or taxonomy calculations will be performed; the pipeline will only upload the data. Study for submission was already registered and provided via --submission_study.' + config_profile_description = 'Tests single MAG submission with all metadata provided in the samplesheet, so the pipeline skips statistics and taxonomy calculation and only performs the upload to ENA. Submits to a pre-existing ENA study passed via --submission_study.' // Input data input = params.pipelines_testdata_base_path + 'seqsubmit/samplesheets/mag_complete_metadata.csv' From 99970b6ddbbbbfe3449fce2ae51dbc180dd9349d Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova <49755902+KateSakharova@users.noreply.github.com> Date: Mon, 27 Jul 2026 15:08:52 +0100 Subject: [PATCH 39/47] Update subworkflows/local/genome_evaluation/main.nf Co-authored-by: Sofia Ochkalova <61541445+ochkalova@users.noreply.github.com> --- subworkflows/local/genome_evaluation/main.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/subworkflows/local/genome_evaluation/main.nf b/subworkflows/local/genome_evaluation/main.nf index 4c477ee..7d78b8d 100644 --- a/subworkflows/local/genome_evaluation/main.nf +++ b/subworkflows/local/genome_evaluation/main.nf @@ -21,8 +21,8 @@ workflow GENOME_EVALUATION { take: ch_fasta // channel: [ val(meta), path(fasta) ] - ch_checkm2_db // val: path to CheckM2 database or null - ch_checkm2_db_download_id // val: CheckM2 database download ID + ch_checkm2_db // val: path to local CheckM2 database or null to trigger automatic download + ch_checkm2_db_download_id // val: CheckM2 database Zenodo download ID main: From 1a322b6c55e04adfd481fd4a76009615f7ee28fa Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova <49755902+KateSakharova@users.noreply.github.com> Date: Mon, 27 Jul 2026 15:09:02 +0100 Subject: [PATCH 40/47] Update conf/test_assembly_complete_metadata.config Co-authored-by: Sofia Ochkalova <61541445+ochkalova@users.noreply.github.com> --- conf/test_assembly_complete_metadata.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test_assembly_complete_metadata.config b/conf/test_assembly_complete_metadata.config index aa8c4d9..801cab7 100644 --- a/conf/test_assembly_complete_metadata.config +++ b/conf/test_assembly_complete_metadata.config @@ -20,7 +20,7 @@ process { params { config_profile_name = 'Test --mode metagenomic_assemblies complete_metadata profile' - config_profile_description = 'Single-case assembly test with complete metadata in samplesheet. No statistics calculations will be performed; the pipeline will only upload the data. Study for submission was already registered and provided via --submission_study.' + config_profile_description = 'Tests single assembly submission with all metadata provided in the samplesheet so the pipeline skips coverage calculation and only performs the upload to ENA. Submits to a pre-existing ENA study passed via --submission_study.' // Input data input = params.pipelines_testdata_base_path + 'seqsubmit/samplesheets/assembly_complete_metadata.csv' From 2a0364cca689580a66a939dec052b52a7e3fb6af Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Mon, 27 Jul 2026 15:38:11 +0100 Subject: [PATCH 41/47] remove nr from main workflow and add to taxonomy subwf --- main.nf | 1 - workflows/genomesubmit.nf | 8 ++++---- 2 files changed, 4 insertions(+), 5 deletions(-) diff --git a/main.nf b/main.nf index 2ba497e..1c9223a 100644 --- a/main.nf +++ b/main.nf @@ -55,7 +55,6 @@ workflow NFCORE_SEQSUBMIT { params.checkm2_db, params.checkm2_db_download_id, params.cat_db, - "nr", // NCBI non-redundant protein database identifier params.centre_name, params.upload_tpa, params.test_upload, diff --git a/workflows/genomesubmit.nf b/workflows/genomesubmit.nf index 2cc74b3..37ce955 100644 --- a/workflows/genomesubmit.nf +++ b/workflows/genomesubmit.nf @@ -46,7 +46,6 @@ workflow GENOMESUBMIT { checkm2_db // val: path to CheckM2 database checkm2_db_download_id // val: CheckM2 database download ID cat_db // val: path to CAT database - cat_db_download_id // val: CAT database download ID centre_name // val: submission centre name upload_tpa // val: upload as TPA (Third Party Annotation) test_upload // val: true for test upload mode @@ -185,9 +184,10 @@ workflow GENOMESUBMIT { ? channel.of( [['id': 'CAT_DB'], file(cat_db)] ) : channel.empty() - def cat_db_id_input = (!cat_db && cat_db_download_id) - ? channel.of( [['id': 'CAT_DB_id'], cat_db_download_id] ) - : channel.empty() + cat_db_download_id = "nr" // NCBI non-redundant protein database identifier + def cat_db_id_input = cat_db + ? channel.empty() + : channel.of( [['id': 'CAT_DB_id'], cat_db_download_id] ) FASTA_CLASSIFY_CATPACK ( RENAME_FASTA_FOR_CATPACK.out.renamed_fasta, // ch_bins From 8af5f5053704ef3e737b002f17d61d2895263ea1 Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova <49755902+KateSakharova@users.noreply.github.com> Date: Mon, 27 Jul 2026 15:39:08 +0100 Subject: [PATCH 42/47] Update docs and schemas for better descriptions Co-authored-by: Sofia Ochkalova <61541445+ochkalova@users.noreply.github.com> --- docs/CONTRIBUTING.md | 2 +- docs/methods.md | 2 +- docs/usage.md | 4 ++-- nextflow_schema.json | 18 +++++++++--------- 4 files changed, 13 insertions(+), 13 deletions(-) diff --git a/docs/CONTRIBUTING.md b/docs/CONTRIBUTING.md index 2ff2980..7330210 100644 --- a/docs/CONTRIBUTING.md +++ b/docs/CONTRIBUTING.md @@ -186,7 +186,7 @@ To ensure consistency and maintainability of the nf-core/seqsubmit pipeline, con ### Code style -The pipeline is organized into three separate workflows `(mode`s-) because each mode has distinct procedures and requirements. Future development should follow the same structure to maintain consistency and readability. +The pipeline is organized into three separate workflows, one for each mode (mags, bins, metagenomic_assemblies, reads), because each data type has distinct submission procedures and metadata requirements. Future development should follow the same structure to maintain consistency and readability. If a new feature extends support for an **existing** data type (for example, _adding support for eukaryotic MAGs_), the corresponding logic should be integrated into the existing workflow rather than creating a new one. diff --git a/docs/methods.md b/docs/methods.md index 567e19a..1504fa1 100644 --- a/docs/methods.md +++ b/docs/methods.md @@ -125,7 +125,7 @@ If `NCBI_lineage` is already present in the input samplesheet, the value is reta Before classification, input FASTA files are normalized to a `.fasta` suffix by `RENAME_FASTA_FOR_CATPACK`. Classification is then run in bin mode (`CAT_pack bins`, followed by `CAT_pack add_names`). -Database input is taken from `--cat_db` when provided; otherwise the workflow uses `nr` database identifier to download and prepare a CATPACK NCBI database. See [CAT_pack documentation](https://github.com/MGXlab/CAT_pack#downloading-preconstructed-database-files). There are two options available: `nr` - NCBI non-redundant protein database and `gtdb` (Genome Taxonomy Database (GTDB) proteins). We use NCBI proteins because ENA supports only NCBI format of taxonomy for submission. The resulting classification table is parsed, and the lineage field is written to `NCBI_lineage`. +Database input is taken from `--cat_db` when provided; otherwise the workflow builds CAT_pack database from NCBI non-redundant protein database (`nr`). See [CAT_pack documentation](https://github.com/MGXlab/CAT_pack#downloading-preconstructed-database-files). There are two options available: `nr` - NCBI non-redundant protein database and `gtdb` (Genome Taxonomy Database (GTDB) proteins). We use NCBI proteins because ENA supports only NCBI format of taxonomy for submission. The resulting classification table is parsed, and the lineage field is written to `NCBI_lineage`. ### Completeness and contamination assessment diff --git a/docs/usage.md b/docs/usage.md index 353a906..ff4ec7c 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -337,7 +337,7 @@ In particular, the CAT_pack database is extremely large and may take a long time - provide the path to a local database with `--checkm2_db`, otherwise the pipeline downloads the version specified with `--checkm2_db_download_id` (by default Zenodo accession `14897628`). - CAT_pack: - - provide the path to a local database (containing `tax/` and `db/` folders or a `tar.gz` archive) with `--cat_db`, otherwise the pipeline constructs NCBI non-redundant protein database. + - provide the path to a local database (containing `tax/` and `db/` folders or a `tar.gz` archive) with `--cat_db`, otherwise the pipeline builds CAT_pack DB from NCBI non-redundant protein database (`nr`). See the [CAT_pack documentation](https://github.com/MGXlab/CAT_pack) and [CheckM2 documentation](https://github.com/chklovski/CheckM2) for more details on usage and creation of databases. @@ -352,7 +352,7 @@ In addition to the [common parameters](#running-the-pipeline): | -------------- | -------- | ---------------------------------------------------------------------------------------------------------------------------------------------- | | `--upload_tpa` | No | Mark assemblies as [third party assemblies (TPA)](#third-party-assemblies-tpa) when required. Default: `false`. | | `--checkm2_db` | No | Path to a local CheckM2 database. If omitted, downloads the version set by `--checkm2_db_download_id`. Default: `14897628` (Zenodo accession). | -| `--cat_db` | No | Path to a local CAT_pack database. If omitted, constructs the NCBI non-redundant protein database. | +| `--cat_db` | No | Path to a local CAT_pack database. If omitted, builds CAT_pack DB from NCBI non-redundant protein database (`nr`). | Example `mags` mode upload to TEST server, run command with docker: diff --git a/nextflow_schema.json b/nextflow_schema.json index d6323b9..e69e8d0 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -261,9 +261,9 @@ }, "upload_tpa": { "type": "boolean", - "description": "Flag to control the type of assembly study (third party assembly (TPA) or not). TPA description: https://www.ncbi.nlm.nih.gov/genbank/tpa/.", - "default": false, - "help": "Use this flag if the study is a third party assembly (TPA). Default: false" + "description": "Mark the assemblies as a Third PArty (TPA)", + "help": "Set to true if the assembly was built from public sequence data (TPA) rather than original primary data. See NCBI TPA guidance: https://www.ncbi.nlm.nih.gov/genbank/tpa/", + "default": false }, "mode": { "type": "string", @@ -280,15 +280,15 @@ }, "is_private": { "type": "boolean", - "description": "Use that flag if you are referring to private data in ENA", "default": false, - "help": "This flag is used during metadata fetch steps. Specify if you are referring to private data (project/reads/assembly). That flag does NOT regulate privacy of your submission data (check --release_date)" + "description": "Use that flag if you are referencing private data accessions in the samplesheet", + "help": "Set to true when the referenced project/reads/assembly are still private in ENA, so metadata is fetched via authenticated access. Does NOT affect the privacy of your own submission, use --release_date for that" }, "release_date": { "type": "string", "pattern": "^\\d{4}-\\d{2}-\\d{2}$", - "description": "Use that flag if you want to keep your data private after submission", - "help": "This flag regulates when the created study for submission will become publically available and indexed on the ENA. If you want to keep your data private until a later date, you need to specify a DATE when it can become public (YYYY-MM-DD, max 2 years from now)" + "description": "Date (YYYY-MM-DD) until which the newly created study stays private in ENA.", + "help": "Sets the release date for the study created by the pipeline. Data stays private until this date, then is automatically released and indexed by ENA. Must be no more than 2 years from today. Omit to release the study publicly immediately." }, "submission_study": { "type": "string", @@ -299,8 +299,8 @@ "type": "string", "format": "file-path", "exists": true, - "description": "Path to study metadata file (JSON, CSV, or TSV) for registering a new ENA study. Required when submission_study is not provided.", - "help_text": "File containing study metadata fields (required: study_title and alias, optional: study_abstract, existing_study_type, etc.). Used by REGISTERSTUDY to create a new study in ENA when no existing submission_study accession is given. ENA documentation: https://ena-docs.readthedocs.io/en/latest/submit/study/programmatic.html", + "description": "Path to a file (JSON, CSV, or TSV) with metadata for registering a new ENA study. Required if --submission_study is not set.", + "help_text": "File containing study metadata fields (required: study_title, alias; optional: study_abstract, existing_study_type, etc.) used by the pipeline to register a new ENA study when no existing --submission_study accession is given. ENA docs: https://ena-docs.readthedocs.io/en/latest/submit/study/programmatic.html", "fa_icon": "fas fa-file-alt" }, "webincli_mode": { From b4a5d4852b580587d14260e2b6e9ff4af5495b8d Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Tue, 28 Jul 2026 13:57:45 +0100 Subject: [PATCH 43/47] aligned docs and schemas --- assets/schema_input_assembly.json | 15 +++-- assets/schema_input_genome.json | 28 ++++---- assets/schema_input_reads.json | 21 +++--- docs/usage.md | 108 +++++++++++++++--------------- nextflow_schema.json | 42 ++++++------ 5 files changed, 108 insertions(+), 106 deletions(-) diff --git a/assets/schema_input_assembly.json b/assets/schema_input_assembly.json index 4a767ca..308ccc1 100644 --- a/assets/schema_input_assembly.json +++ b/assets/schema_input_assembly.json @@ -10,7 +10,7 @@ "id": { "type": "string", "pattern": "^\\S+$", - "description": "Unique assembly identifier", + "description": "A unique identifier for this data entry. Must be globally unique within the input dataset", "errorMessage": "ID must be provided and cannot contain spaces", "meta": ["id"] }, @@ -19,7 +19,7 @@ "format": "file-path", "exists": true, "pattern": "^([\\S\\s]*\\/)?[^\\s\\/]+\\.(fa|fasta|fna)\\.gz$", - "errorMessage": "FASTA file must be provided and have extension '.fa.gz', '.fasta.gz', '.fna.gz'", + "errorMessage": "Path to assembly contigs in FASTA format compressed with gzip", "description": "Metagenomic assembly FASTA file" }, "fastq_1": { @@ -35,8 +35,9 @@ "maxLength": 0 } ], + "default": null, "errorMessage": "FASTQ file must have extension '.fq' or '.fastq' (optionally gzipped)", - "description": "Forward reads if paired-end or single-end reads FASTQ file" + "description": "Path to single-end/paired-end forward reads in FASTQ format (optionally gzipped). Required if coverage is not provided" }, "fastq_2": { "anyOf": [ @@ -53,7 +54,7 @@ ], "default": null, "errorMessage": "FASTQ file for reverse reads must have extension '.fq' or '.fastq' (optionally gzipped)", - "description": "Reverse reads FASTQ file if paired-end. Leave empty for single-end reads" + "description": "Path to reverse reads in FASTQ format for paired-end data (optionally gzipped). Leave empty for single-end reads" }, "coverage": { "anyOf": [ @@ -68,14 +69,14 @@ ], "default": null, "errorMessage": "Coverage must be a positive number or empty", - "description": "Estimated value of assembly coverage", + "description": "Estimated sequencing depth of the assembly (optional). If this value is missing, it is computed automatically during pipeline execution when reads are provided", "meta": ["coverage"] }, "run_accession": { "type": "string", "pattern": "^\\S+$", "errorMessage": "Accession must be provided and cannot contain spaces", - "description": "Accession of the run used to generate the assembly", + "description": "ENA run accession for the reads used to generate the metagenomic assembly. Reads must already be submitted to ENA", "meta": ["run_accession"] }, "assembler": { @@ -89,7 +90,7 @@ "anyOf": [{ "type": "string" }, { "type": "number" }], "pattern": "^\\S+$", "errorMessage": "Assembler version must be provided and cannot contain spaces", - "description": "Version of the assembler software used to generate the assembly, e.g 1.0.0", + "description": "Version of the assembler software used to generate the assembly, e.g. 1.0.0", "meta": ["assembler_version"] } }, diff --git a/assets/schema_input_genome.json b/assets/schema_input_genome.json index 28c4d69..2187d86 100644 --- a/assets/schema_input_genome.json +++ b/assets/schema_input_genome.json @@ -10,7 +10,7 @@ "id": { "type": "string", "pattern": "^\\S+$", - "description": "Unique mag/bin identifier", + "description": "A unique identifier for this data entry. Must be globally unique within the input dataset", "errorMessage": "ID must be provided and cannot contain spaces", "meta": ["id"] }, @@ -19,12 +19,12 @@ "format": "file-path", "exists": true, "pattern": "^([\\S\\s]*\\/)?[^\\s\\/]+\\.(fa|fasta|fna)\\.gz$", - "errorMessage": "FASTA file for sequences 1 must be provided, cannot contain spaces and must have extension '.fa.gz', '.fasta.gz', or '.fna.gz'", + "errorMessage": "Path to MAG/bin sequences in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. File should contain at least one sequence", "description": "MAG/bin sequence file" }, "accession": { "type": "string", - "description": "Run or assembly ENA accession", + "description": "ENA accession of the run or metagenomic assembly used to generate the MAG/bin", "meta": ["accession"] }, "fastq_1": { @@ -42,7 +42,7 @@ ], "default": null, "errorMessage": "FASTQ file must have extension '.fq' or '.fastq' (optionally gzipped)", - "description": "Forward reads if paired-end or single-end reads FASTQ file" + "description": "Path to single-end/paired-end forward reads (in FASTQ format, optionally gzipped) used to generate the source metagenomic assembly. Required if genome_coverage is not provided" }, "fastq_2": { "anyOf": [ @@ -59,7 +59,7 @@ ], "default": null, "errorMessage": "FASTQ file for reverse reads must have extension '.fq' or '.fastq' (optionally gzipped)", - "description": "Reverse reads FASTQ file if paired-end. Leave empty for single-end reads" + "description": "Path to reverse reads in FASTQ format for paired-end data (optionally gzipped) used to generate the source metagenomic assembly. Leave empty for single-end reads" }, "assembly_software": { "type": "string", @@ -68,18 +68,18 @@ }, "binning_software": { "type": "string", - "description": "Tool name and version that was used to bin data, e.g. MetaBAT2, VAMB, MaxBin2, CONCOCT, SemiBin2, COMEBin, etc", + "description": "Tool name and version that was used to bin data, e.g. MetaBAT2_v1.0, VAMB_2.0, MaxBin2_v1, CONCOCT_v3.0, SemiBin2_v1, COMEBin_v2.3.4, etc", "meta": ["binning_software"] }, "binning_parameters": { "type": "string", - "description": "Arguments used to bin data different from default, e.g. -min_contig_length 1000", + "description": "Arguments used to bin data different from default, e.g. -min_contig_length 1000, otherwise put 'default'", "meta": ["binning_parameters"] }, "stats_generation_software": { "type": "string", "default": null, - "description": "Tool(s) used to estimate completeness and contamination", + "description": "Tool, including version, that was used to calculate completeness and contamination, e.g. CheckM2_v1.0.0", "meta": ["stats_generation_software"] }, "completeness": { @@ -95,7 +95,7 @@ } ], "default": null, - "description": "MAG/bin completeness score: the ratio of observed single-copy marker genes to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined", + "description": "MAG/bin completeness score: the 0<= ratio <= 100 of observed single-copy marker genes to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined", "meta": ["completeness"] }, "contamination": { @@ -111,7 +111,7 @@ } ], "default": null, - "description": "MAG/bin contamination score: the ratio of observed single-copy marker genes in ≥2 copies to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined", + "description": "MAG/bin contamination score: the 0<= ratio <= 100 of observed single-copy marker genes in ≥2 copies to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined", "meta": ["contamination"] }, "genome_coverage": { @@ -126,17 +126,17 @@ } ], "default": null, - "description": "MAG/bin coverage value", + "description": "Estimated average sequencing depth (>0) across the genome. If the value is missing, it is computed automatically during pipeline execution when reads are provided", "meta": ["genome_coverage"] }, "metagenome": { "type": "string", - "description": "ENA accepted metagenome name. It needs to be listed in the taxonomy tree https://www.ebi.ac.uk/ena/browser/view/408169?show=tax-tree (you might need to press \"Tax tree - Show\" in the right most section of the page). Full list can also be found https://github.com/EBI-Metagenomics/genome_uploader/blob/main/genomeuploader/constants.py#L22", + "description": "Registered metagenome taxonomic identifier or name that matches an existing ENA taxonomy entry. It needs to be listed in the taxonomy tree https://www.ebi.ac.uk/ena/browser/view/408169?show=tax-tree (you might need to press \"Tax tree - Show\" in the right most section of the page). Full list can also be found https://github.com/EBI-Metagenomics/genome_uploader/blob/main/genomeuploader/constants.py#L22", "meta": ["metagenome"] }, "co-assembly": { "type": "boolean", - "description": "'true' if data was co-assembled, 'false' otherwise", + "description": "Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: 'true' or 'false'", "meta": ["co_assembly"] }, "broad_environment": { @@ -165,7 +165,7 @@ } ], "default": null, - "description": "Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: true/false", + "description": "Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: 'true' or 'false'", "meta": ["RNA_presence"] }, "NCBI_lineage": { diff --git a/assets/schema_input_reads.json b/assets/schema_input_reads.json index e39c89e..64931d3 100644 --- a/assets/schema_input_reads.json +++ b/assets/schema_input_reads.json @@ -12,13 +12,13 @@ "pattern": "^\\S+$", "errorMessage": "ID must be provided and cannot contain spaces", "meta": ["id"], - "description": "Unique experiment/run name" + "description": "Unique identifier of this particular data entry. Used as an experiment name" }, "sample_accession": { "type": "string", "pattern": "^\\S+$", "errorMessage": "Sample accession must be provided and cannot contain spaces", - "description": "ENA sample accession of the sample used to generate the reads", + "description": "ENA sample accession (starting with SAMEA) of the sample used to generate raw reads", "meta": ["sample_accession"] }, "fastq_1": { @@ -27,7 +27,7 @@ "exists": true, "pattern": "^\\S+\\.(fq|fastq)(\\.gz)?$", "errorMessage": "FASTQ file must have extension '.fq' or '.fastq' (optionally gzipped)", - "description": "Forward reads FASTQ file (single-end or paired-end)" + "description": "Path to single-end/paired-end forward reads in FASTQ format (optionally gzipped)" }, "fastq_2": { "anyOf": [ @@ -44,7 +44,7 @@ ], "default": null, "errorMessage": "FASTQ file for reverse reads must have extension '.fq' or '.fastq' (optionally gzipped)", - "description": "Reverse reads FASTQ file if paired-end. Leave empty for single-end reads" + "description": "Path to reverse reads in FASTQ format for paired-end data (optionally gzipped). Leave empty for single-end reads" }, "platform": { "type": "string", @@ -65,14 +65,14 @@ "ULTIMA", "VELA_DIAGNOSTICS" ], - "description": "Sequencing platform. Must be one of the ENA controlled vocabulary values listed in the enum.", + "description": "Sequencing platform. Must be one of the ENA controlled vocabulary values listed in https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation.", "meta": ["platform"] }, "instrument": { "type": "string", "pattern": "^[^\\n]+$", "errorMessage": "Instrument must be provided and cannot span multiple lines", - "description": "Sequencer model (e.g., 'Illumina HiSeq 2000', 'PacBio Sequel')", + "description": "Sequencer model (e.g., 'Illumina HiSeq 2000', 'PacBio Sequel'). Must be one of the allowed ENA controlled vocabulary values listed in https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#permitted-values-for-instrument", "meta": ["instrument"] }, "library_source": { @@ -88,7 +88,7 @@ "VIRAL RNA", "OTHER" ], - "description": "Library source. Must be one of the ENA controlled vocabulary values listed in the enum.", + "description": "Library source type. Must be one of the allowed ENA controlled vocabulary values listed in https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#permitted-values-for-library-source", "meta": ["library_source"] }, "library_selection": { @@ -126,7 +126,7 @@ "other", "unspecified" ], - "description": "Library selection. Must be one of the allowed ENA controlled vocabulary values (e.g. PCR, ChIP-Seq, other, etc).", + "description": "Library selection. Must be one of the allowed ENA controlled vocabulary values listed in https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#permitted-values-for-library-selection", "meta": ["library_selection"] }, "library_strategy": { @@ -174,7 +174,7 @@ "Tethered Chromatin Conformation Capture", "OTHER" ], - "description": "Library strategy. Must be one of the ENA controlled vocabulary values listed in the enum.", + "description": "Library strategy. Must be one of the ENA controlled vocabulary values listed in https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#permitted-values-for-library-strategy", "meta": ["library_strategy"] }, "insert_size": { @@ -188,8 +188,9 @@ "maxLength": 0 } ], + "default": null, "errorMessage": "Insert size must be a positive number or empty", - "description": "Fragment/insert size for paired-end reads (optional)", + "description": "Fragment/insert size for paired-end reads (e.g., 500 for 500 bp inserts). Skip if not applicable.", "meta": ["insert_size"] }, "library_name": { diff --git a/docs/usage.md b/docs/usage.md index ff4ec7c..a82cec4 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -161,20 +161,20 @@ illumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLU pacbio_run_001,SAMEA7654321,data/pacbio_reads.fastq.gz,,PACBIO_SMRT,PacBio Sequel,GENOMIC,RANDOM,WGS,,PacBio_library_002,Long-read sequencing ``` -| Column | Required | Description | -| ------------------- | -------- | --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | -| `id` | Yes | Unique identifier of this particular data entry. Used as an experiment name. | -| `sample_accession` | Yes | ENA sample accession (starting with SAMEA) of the sample used to generate raw reads. | -| `fastq_1` | Yes | Path to forward reads in FASTQ format (optionally gzipped). | -| `fastq_2` | No | Path to reverse reads for paired-end data. Leave empty for single-end reads. | -| `platform` | Yes | Sequencing platform. Supported values: `ILLUMINA`, `PACBIO_SMRT`, `OXFORD_NANOPORE`, `ION_TORRENT`, `CAPILLARY`, `DNBSEQ`, `ELEMENT`, `GENAPSYS`, `GENEMIND`, `HELICOS`, `LS454`, `BGISEQ`, `ULTIMA`, `VELA_DIAGNOSTICS`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | -| `instrument` | Yes | Sequencer model, e.g. "Illumina HiSeq 2000", "PacBio Sequel", "MinION". See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#permitted-values-for-instrument). | -| `library_source` | Yes | Library source type. Options: `GENOMIC`, `METAGENOMIC`, `TRANSCRIPTOMIC`, `METAGENOMIC SINGLE CELL`, `TRANSCRIPTOMIC SINGLE CELL`, `SYNTHETIC`, `VIRAL RNA`, `OTHER`. | -| `library_selection` | Yes | Library selection method. Options: `RANDOM`, `PCR`, `RANDOM PCR`, `RT-PCR`, `MF`, `cDNA`, `cDNA_randomPriming`, `cDNA_oligo_dT`, `PolyA`, `Inverse rRNA`, `ChIP`, `MNase`, `DNase`, `Hybrid Selection`, etc. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | -| `library_strategy` | Yes | Library strategy. Options: `WGS`, `WGA`, `WXS`, `RNA-Seq`, `miRNA-Seq`, `ncRNA-Seq`, `EST`, `Hi-C`, `ATAC-seq`, `WCS`, `RAD-Seq`, `CLONE`, `AMPLICON`, `POOLCLONE`, `etc`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation) for complete list. | -| `insert_size` | No | Fragment/insert size for paired-end reads (e.g., 500 for 500 bp inserts). Leave empty if not applicable. | -| `library_name` | No | Descriptive library name (optional). | -| `description` | No | Free-text description of the experiment (optional). | +| Column | Required | Description | +| ------------------- | -------- | ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `id` | Yes | Unique identifier of this particular data entry. Used as an experiment name. | +| `sample_accession` | Yes | ENA sample accession (starting with SAMEA) of the sample used to generate raw reads. | +| `fastq_1` | Yes | Path to single-end/paired-end forward reads in FASTQ format (optionally gzipped). | +| `fastq_2` | No | Path to reverse reads in FASTQ format for paired-end data (optionally gzipped). Leave empty for single-end reads. | +| `platform` | Yes | Sequencing platform. Must be one of the ENA controlled vocabulary values listed in [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation). Supported values: `ILLUMINA`, `PACBIO_SMRT`, `OXFORD_NANOPORE`, `ION_TORRENT`, `CAPILLARY`, `DNBSEQ`, `ELEMENT`, `GENAPSYS`, `GENEMIND`, `HELICOS`, `LS454`, `BGISEQ`, `ULTIMA`, `VELA_DIAGNOSTICS`. | +| `instrument` | Yes | Sequencer model, e.g. "Illumina HiSeq 2000", "PacBio Sequel", "MinION". See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#permitted-values-for-instrument). | +| `library_source` | Yes | Library source type. Options: `GENOMIC`, `METAGENOMIC`, `TRANSCRIPTOMIC`, `METAGENOMIC SINGLE CELL`, `TRANSCRIPTOMIC SINGLE CELL`, `SYNTHETIC`, `VIRAL RNA`, `OTHER`. ENA [documantation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#permitted-values-for-library-source). | +| `library_selection` | Yes | Library selection method. Options: `RANDOM`, `PCR`, `RANDOM PCR`, `RT-PCR`, `MF`, `cDNA`, `cDNA_randomPriming`, `cDNA_oligo_dT`, `PolyA`, `Inverse rRNA`, `ChIP`, `MNase`, `DNase`, `Hybrid Selection`, etc. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#permitted-values-for-library-selection) for complete list. | +| `library_strategy` | Yes | Library strategy. Options: `WGS`, `WGA`, `WXS`, `RNA-Seq`, `miRNA-Seq`, `ncRNA-Seq`, `EST`, `Hi-C`, `ATAC-seq`, `WCS`, `RAD-Seq`, `CLONE`, `AMPLICON`, `POOLCLONE`, `etc`. See [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#permitted-values-for-library-strategy) for complete list. | +| `insert_size` | No | Fragment/insert size for paired-end reads (e.g., 500 for 500 bp inserts). Leave empty if not applicable. | +| `library_name` | No | Descriptive library name (optional). | +| `description` | No | Free-text description of the experiment (optional). | ### `reads` parameters and example @@ -223,12 +223,12 @@ assembly_002,data/assembly_002.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9 | ------------------- | ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------- | | `id` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset. | | `fasta` | Yes | Path to assembly contigs in FASTA format compressed with `gzip`. | -| `fastq_1` | Conditional | Path to the read file in FASTQ format used to generate the metagenomic assembly. Required if `coverage` is not provided. | +| `fastq_1` | Conditional | Path to single-end/paired-end forward reads in FASTQ format (optionally gzipped). Required if `coverage` is not provided. | | `fastq_2` | No | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | | `coverage` | Conditional | Estimated sequencing depth of the assembly. If this value is missing, it is computed automatically during pipeline execution when reads are provided. | | `run_accession` | Yes | ENA run accession for the reads used to generate the metagenomic assembly. Reads must already be submitted to ENA. | -| `assembler` | Yes | Name of the assembler software used to generate the assembly. | -| `assembler_version` | Yes | Version of the assembler software used to generate the assembly. | +| `assembler` | Yes | Name of the assembler software used to generate the assembly, e.g. MEGAHIT, SPAdes, metaSPAdes, Flye, etc. | +| `assembler_version` | Yes | Version of the assembler software used to generate the assembly, e.g. 1.0.0. | Provide either read files (`fastq_1`, optionally `fastq_2`) or a `coverage` value for each row. If `coverage` is missing and reads are provided, the workflow calculates average coverage with CoverM. See the [Methods documentation](docs/methods.md) for more information on how coverage is calculated. @@ -297,27 +297,27 @@ id,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_pa mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default,CheckM2 1.0.1,92.81,1.09,66.04,sediment metagenome,false,marine,cable bacteria,marine sediment,false,d__Bacteria;p__Proteobacteria;s__ ``` -| Column | Required | Description | -| --------------------------- | ----------- | ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | -| `id` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset. | -| `fasta` | Yes | Path to MAG/bin contigs in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. | -| `accession` | Yes | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | -| `fastq_1` | Conditional | Path to the read file in FASTQ format used to generate the source metagenomic assembly. Required if `genome_coverage` is not provided. | -| `fastq_2` | No | Path to the second read file in FASTQ format for paired-end data used to generate the source metagenomic assembly. Leave empty for single-end reads. | -| `assembly_software` | Yes | Tool name and version that were used to generate the source metagenomic assembly. | -| `binning_software` | Yes | Binning tool, including version, that was used to generate the bins. | -| `binning_parameters` | Yes | Arguments that were used during binning. | -| `stats_generation_software` | No | Tool, including version, that was used to calculate completeness and contamination. | -| `completeness` | No | Genome completeness value. | -| `contamination` | No | Genome contamination value. | -| `genome_coverage` | Conditional | Estimated average sequencing depth across the genome. If the value is missing, it is computed automatically during pipeline execution when reads are provided. | -| `metagenome` | Yes | Registered metagenome taxonomic identifier or name that matches an existing ENA taxonomy entry. For more details see https://ena-docs.readthedocs.io/en/latest/faq/taxonomy.html | -| `co-assembly` | Yes | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: true or false. | -| `broad_environment` | Yes | Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428) | -| `local_environment` | Yes | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". | -| `environmental_medium` | Yes | Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). | -| `RNA_presence` | No | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: true or false. | -| `NCBI_lineage` | No | NCBI taxonomy lineage of the genome. Can be composted of either numerical IDs or official NCBI taxon names separated by ";". | +| Column | Required | Description | +| --------------------------- | ----------- | -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `id` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset. | +| `fasta` | Yes | Path to MAG/bin sequences in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. File should contain at least one sequence. | +| `accession` | Yes | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | +| `fastq_1` | Conditional | Path to single-end/paired-end forward reads in FASTQ format (optionally gzipped) used to generate the source metagenomic assembly. Required if `genome_coverage` is not provided. | +| `fastq_2` | No | Path to reverse reads in FASTQ format for paired-end data (optionally gzipped) used to generate the source metagenomic assembly. Leave empty for single-end reads. | +| `assembly_software` | Yes | Tool name and version that were used to generate the source metagenomic assembly, e.g. MEGAHIT_v1.0, SPAdes_v4.0.0, metaSPAdes_v3.15.0. | +| `binning_software` | Yes | Tool name and version that was used to bin data, e.g. MetaBAT2_v1.0, VAMB_2.0, MaxBin2_v1, CONCOCT_v3.0, SemiBin2_v1, COMEBin_v2.3.4, etc. | +| `binning_parameters` | Yes | Arguments used to bin data different from default, e.g. -min_contig_length 1000, otherwise put 'default'. | +| `stats_generation_software` | No | Tool, including version, that was used to calculate completeness and contamination, e.g. CheckM2_v1.0.0. | +| `completeness` | No | MAG/bin completeness score: the 0<= ratio <= 100 of observed single-copy marker genes to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined. | +| `contamination` | No | MAG/bin contamination score: the 0<= ratio <= 100 of observed single-copy marker genes in ≥2 copies to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined. | +| `genome_coverage` | Conditional | Estimated average sequencing depth (>0) across the genome. If the value is missing, it is computed automatically during pipeline execution when reads are provided. | +| `metagenome` | Yes | Registered metagenome taxonomic identifier or name that matches an existing ENA taxonomy entry. It needs to be listed in the taxonomy tree https://www.ebi.ac.uk/ena/browser/view/408169?show=tax-tree (you might need to press "Tax tree - Show" in the right most section of the page). Full list can also be found https://github.com/EBI-Metagenomics/genome_uploader/blob/main/genomeuploader/constants.py#L22. | +| `co-assembly` | Yes | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: true or false. | +| `broad_environment` | Yes | Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428). Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS. | +| `local_environment` | Yes | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS. | +| `environmental_medium` | Yes | Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS. | +| `RNA_presence` | No | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: true or false. | +| `NCBI_lineage` | No | Full NCBI lineage - format: x;y;z. For example, the lineage for E. coli can be: 'Bacteria;Pseudomonadati;Pseudomonadota;Gammaproteobacteria;Enterobacterales;Enterobacteriaceae;Escherichia' or '2;1224;1236;91347;543;561;562'. For more info check https://www.ncbi.nlm.nih.gov/datasets/docs/v2/data-processing/taxonomy-processing/taxonomy/. | > [!NOTE] > More information about environment tags can be found at checklists [ERC000050](https://www.ebi.ac.uk/ena/browser/view/ERC000050) for bins and [ERC000047](https://www.ebi.ac.uk/ena/browser/view/ERC000047) for MAGs under the field names "broad-scale environmental context", "local environmental context", and "environmental medium". @@ -348,11 +348,11 @@ Databases created/downloaded by the pipeline are published under: In addition to the [common parameters](#running-the-pipeline): -| Parameter | Required | Description | -| -------------- | -------- | ---------------------------------------------------------------------------------------------------------------------------------------------- | -| `--upload_tpa` | No | Mark assemblies as [third party assemblies (TPA)](#third-party-assemblies-tpa) when required. Default: `false`. | -| `--checkm2_db` | No | Path to a local CheckM2 database. If omitted, downloads the version set by `--checkm2_db_download_id`. Default: `14897628` (Zenodo accession). | -| `--cat_db` | No | Path to a local CAT_pack database. If omitted, builds CAT_pack DB from NCBI non-redundant protein database (`nr`). | +| Parameter | Required | Description | +| -------------- | -------- | ------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `--upload_tpa` | No | Mark assemblies as [third party assemblies (TPA)](#third-party-assemblies-tpa) when required. Default: `false`. | +| `--checkm2_db` | No | Path to a pre-downloaded CheckM2 database. If omitted, downloads the version set by `--checkm2_db_download_id`. Default: `14897628` (Zenodo accession). | +| `--cat_db` | No | Path to a local CAT_pack database. If omitted, builds CAT_pack DB from NCBI non-redundant protein database (`nr`). | Example `mags` mode upload to TEST server, run command with docker: @@ -406,17 +406,17 @@ nextflow run nf-core/seqsubmit \ Parameters common to all modes: -| Parameter | Required | Description | -| ------------------------------------------ | ------------ | -------------------------------------------------------------------------------------------------------------------- | -| `--mode` | Yes | Type of the data to be submitted. Options: `mags`, `bins`, `metagenomic_assemblies`, `reads`. | -| `--input` | Yes | Path to the samplesheet describing the data to submit. | -| `--outdir` | Yes | Path to the output directory for pipeline results. | -| `--submission_study` OR `--study_metadata` | Yes (one of) | ENA study accession (PRJ/ERP) to submit the data to OR metadata file in JSON/TSV/CSV format to register a new study. | -| `--centre_name` | Yes | Name of the submitter's organisation. | -| `--test_upload` | No | Submit to the ENA TEST server instead of the LIVE server. Default: `true`. | -| `--webincli_mode` | No | Webin-CLI mode for ENA interaction: `submit` uploads data, `validate` performs validation only. Default: `submit`. | -| `--is_private` | No | Use if you are referencing private data accessions in your submission. Default: `false`. | -| `--release_date` | No | Use if you want to keep your data private after submission until a particular date. | +| Parameter | Required | Description | +| ------------------------------------------ | ------------ | ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `--mode` | Yes | Type of the data to be submitted. Options: `mags`, `bins`, `metagenomic_assemblies`, `reads`. | +| `--input` | Yes | Path to samplesheet describing the data to be submitted (supported formats: csv, tsv, yaml, yml, json). Columns/fields depend on the pipeline mode. See [usage docs](https://nf-co.re/seqsubmit/usage#samplesheet-input). | +| `--outdir` | Yes | Path to the output directory for pipeline results. You have to use absolute paths to storage on Cloud infrastructure. | +| `--submission_study` OR `--study_metadata` | Yes (one of) | ENA study accession (PRJ/ERP) to submit the data to OR metadata file in JSON/TSV/CSV format to register a new study. | +| `--centre_name` | Yes | Name of the submitter's organisation (mandatory for broker accounts). | +| `--test_upload` | No | Submit to the ENA TEST server instead of the LIVE server. Use that flag for development purposes or for validation of your submissions. Submission would be done to TEST ENA server instead of LIVE server. TEST server holds data 24h and removes afterwords. LIVE server does not have option to modify your submission, you can only suppress data and re-upload. Default: `true`. | +| `--webincli_mode` | No | Webin-CLI mode for ENA interaction: `submit` uploads data, `validate` performs validation only. `validate` mode checks correctness of manifest files, data access and validity. `submit` mode does validation and push data to ENA server. Default: `submit`. | +| `--is_private` | No | Use if you are referencing private data accessions in your submission. Set to true when the referenced project/reads/assembly are still private in ENA, so metadata is fetched via authenticated access. Does NOT affect the privacy of your own submission, use --release_date for that. Default: `false`. | +| `--release_date` | No | Use if you want to keep your data private in ENA after submission until a particular date (format: YYYY-MM-DD). Data stays private until this date, then is automatically released and indexed by ENA. Must be no more than 2 years from today. Omit to release the study publicly immediately. | Each mode also has additional parameters and example commands — see its own "`` parameters and example" subsection above. diff --git a/nextflow_schema.json b/nextflow_schema.json index e69e8d0..70dce8f 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -18,8 +18,8 @@ "exists": true, "mimetype": "text/csv", "pattern": "^\\S+\\.(csv|tsv|yaml|yml|json)$", - "description": "Path to comma-separated file describing the data to be submitted. Format depends on the pipeline mode (mags/bins/metagenomic_assemblies/reads).", - "help_text": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with a set of columns depending on the type of data being submitted. See [usage docs](https://nf-co.re/seqsubmit/usage#samplesheet-input).", + "description": "Path to samplesheet describing the data to be submitted (supported formats: csv, tsv, yaml, yml, json). Columns/fields depend on the pipeline mode", + "help": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a file in supported format with a set of columns depending on the type of data being submitted. See [usage docs](https://nf-co.re/seqsubmit/usage#samplesheet-input).", "fa_icon": "fas fa-file-csv" }, "outdir": { @@ -32,7 +32,7 @@ "type": "string", "description": "Email address for completion summary.", "fa_icon": "fas fa-envelope", - "help_text": "Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (`~/.nextflow/config`) then you don't need to specify this on the command line for every run.", + "help": "Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (`~/.nextflow/config`) then you don't need to specify this on the command line for every run.", "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$" }, "multiqc_title": { @@ -47,7 +47,7 @@ "type": "object", "fa_icon": "fas fa-university", "description": "Parameters affecting a decision about presence/absence of tRNA and rRNA genes (RNA_presence) in a genome", - "help_text": "The value of RNA_presence is based on the number of rRNA and tRNA. If at least 18 tRNAs are detected and 16S, 23S, and 5S rRNA subunits pass the recovery threshold - RNA_presence is True, otherwise - False. The resulting value is a decisive factor affecting MISAG/MIMAG assembly quality classification of a genome.", + "help": "The value of RNA_presence is based on the number of rRNA and tRNA. If at least 18 tRNAs are detected and 16S, 23S, and 5S rRNA subunits pass the recovery threshold - RNA_presence is True, otherwise - False. The resulting value is a decisive factor affecting MISAG/MIMAG assembly quality classification of a genome.", "properties": { "rrna_limit": { "type": "number", @@ -68,7 +68,7 @@ "type": "object", "fa_icon": "fas fa-university", "description": "CAT_pack required data", - "help_text": "Provide a path to pre-downloaded DB or DB id for automatic download", + "help": "Provide a path to pre-downloaded DB or DB id for automatic download", "properties": { "cat_db": { "format": "file-path", @@ -83,16 +83,16 @@ "type": "object", "fa_icon": "fas fa-university", "description": "Parameters used to run Checkm2", - "help_text": "These options are used to run Checkm2 that will calculate completeness and contamination of genome if those were not provided as input", + "help": "These options are used to run Checkm2 that will calculate completeness and contamination of genome if those were not provided as input", "properties": { "checkm2_db": { "format": "file-path", - "description": "Path to pre-downloaded checkm2 database", + "description": "Path to pre-downloaded CheckM2 database. If omitted, downloads the version set by `--checkm2_db_download_id`", "fa_icon": "fas fa-users-cog" }, "checkm2_db_download_id": { "type": "integer", - "description": "Zenodo ID for checkm2 database download", + "description": "Zenodo ID for CheckM2 database download", "default": 14897628, "fa_icon": "fas fa-users-cog" } @@ -103,7 +103,7 @@ "type": "object", "fa_icon": "fas fa-university", "description": "Parameters used to describe centralised config profiles. These should not be edited.", - "help_text": "The centralised nf-core configuration profiles use a handful of pipeline parameters to describe themselves. This information is then printed to the Nextflow log when you run a pipeline. You should not need to change these values when you run a pipeline.", + "help": "The centralised nf-core configuration profiles use a handful of pipeline parameters to describe themselves. This information is then printed to the Nextflow log when you run a pipeline. You should not need to change these values when you run a pipeline.", "properties": { "custom_config_version": { "type": "string", @@ -117,7 +117,7 @@ "description": "Base directory for Institutional configs.", "default": "https://raw.githubusercontent.com/nf-core/configs/master", "hidden": true, - "help_text": "If you're running offline, Nextflow will not be able to fetch the institutional config files from the internet. If you don't need them, then this is not a problem. If you do need them, you should download the files from the repo and tell Nextflow where to find them with this parameter.", + "help": "If you're running offline, Nextflow will not be able to fetch the institutional config files from the internet. If you don't need them, then this is not a problem. If you do need them, you should download the files from the repo and tell Nextflow where to find them with this parameter.", "fa_icon": "fas fa-users-cog" }, "config_profile_name": { @@ -151,7 +151,7 @@ "type": "object", "fa_icon": "fas fa-file-import", "description": "Less common options for the pipeline, typically set in a config file.", - "help_text": "These options are common to all nf-core pipelines and allow you to customise some of the core preferences for how the pipeline runs.\n\nTypically these options would be set in a Nextflow config file loaded for all pipeline runs, such as `~/.nextflow/config`.", + "help": "These options are common to all nf-core pipelines and allow you to customise some of the core preferences for how the pipeline runs.\n\nTypically these options would be set in a Nextflow config file loaded for all pipeline runs, such as `~/.nextflow/config`.", "properties": { "version": { "type": "boolean", @@ -163,7 +163,7 @@ "type": "string", "default": "copy", "description": "Method used to save pipeline results to output directory.", - "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", + "help": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", "fa_icon": "fas fa-copy", "enum": ["symlink", "rellink", "link", "copy", "copyNoFollow", "move"], "hidden": true @@ -173,7 +173,7 @@ "description": "Email address for completion summary, only when pipeline fails.", "fa_icon": "fas fa-exclamation-triangle", "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$", - "help_text": "An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully.", + "help": "An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully.", "hidden": true }, "plaintext_email": { @@ -256,7 +256,7 @@ "properties": { "centre_name": { "type": "string", - "description": "The center name of the submitter (mandatory for broker accounts).", + "description": "Name of the submitter's organisation (mandatory for broker accounts).", "help": "ena-webin-cli argument -centerName: the center name of the submitter (mandatory for broker accounts)." }, "upload_tpa": { @@ -268,13 +268,13 @@ "mode": { "type": "string", "default": null, - "description": "Type of upload", - "help_text": "Different types of data require specific upload steps. That mode controls what upload workflow to run depending on type of data (mags/bins/metagenomic_assemblies/reads)", + "description": "Type of the data to be submitted", + "help": "Different types of data require specific upload steps. That mode controls what upload workflow to run depending on type of data (mags/bins/metagenomic_assemblies/reads)", "enum": ["mags", "bins", "metagenomic_assemblies", "reads"] }, "test_upload": { "type": "boolean", - "description": "Upload to TEST server instead of LIVE", + "description": "Upload to ENA TEST server instead of LIVE server", "default": true, "help": "Use that flag for development purposes or for validation of your submissions. Submission would be done to TEST ENA server instead of LIVE server. TEST server holds data 24h and removes afterwords. LIVE server does not have option to modify your submission, you can only suppress data and re-upload" }, @@ -293,22 +293,22 @@ "submission_study": { "type": "string", "description": "ENA study accession (PRJ/ERP) to submit the data to", - "help_text": "Current implementation of pipeline requires to pre-register ENA project (PRJ/ERP) where you want to upload data to. Documentation how to register study: https://ena-docs.readthedocs.io/en/latest/submit/study.html" + "help": "Current implementation of pipeline requires to pre-register ENA project (PRJ/ERP) where you want to upload data to. Documentation how to register study: https://ena-docs.readthedocs.io/en/latest/submit/study.html" }, "study_metadata": { "type": "string", "format": "file-path", "exists": true, "description": "Path to a file (JSON, CSV, or TSV) with metadata for registering a new ENA study. Required if --submission_study is not set.", - "help_text": "File containing study metadata fields (required: study_title, alias; optional: study_abstract, existing_study_type, etc.) used by the pipeline to register a new ENA study when no existing --submission_study accession is given. ENA docs: https://ena-docs.readthedocs.io/en/latest/submit/study/programmatic.html", + "help": "File containing study metadata fields (required: study_title, alias; optional: study_abstract, existing_study_type, etc.) used by the pipeline to register a new ENA study when no existing --submission_study accession is given. ENA docs: https://ena-docs.readthedocs.io/en/latest/submit/study/programmatic.html", "fa_icon": "fas fa-file-alt" }, "webincli_mode": { "type": "string", - "description": "Webin-CLI mode of operation", + "description": "Webin-CLI mode for ENA interaction: `submit` uploads data, `validate` performs validation only.", "default": "submit", "enum": ["submit", "validate"], - "help": "Controls ENA Webin-CLI mode. Use `submit` to upload data or `validate` to run validation only. Default: submit." + "help": "Controls ENA Webin-CLI mode. `validate` mode checks correctness of manifest files, data access and validity. `submit` mode does validation and push data to ENA server. Default: submit." } } } From 68a9c22553de42a2b77547e9e866b728263cd579 Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Thu, 30 Jul 2026 14:07:00 +0100 Subject: [PATCH 44/47] docs polishing --- assets/schema_input_assembly.json | 4 ++-- assets/schema_input_genome.json | 10 ++++---- assets/schema_input_reads.json | 2 +- docs/usage.md | 8 +++---- nextflow_schema.json | 40 +++++++++++++++---------------- ro-crate-metadata.json | 2 +- 6 files changed, 33 insertions(+), 33 deletions(-) diff --git a/assets/schema_input_assembly.json b/assets/schema_input_assembly.json index 308ccc1..f6a8593 100644 --- a/assets/schema_input_assembly.json +++ b/assets/schema_input_assembly.json @@ -19,8 +19,8 @@ "format": "file-path", "exists": true, "pattern": "^([\\S\\s]*\\/)?[^\\s\\/]+\\.(fa|fasta|fna)\\.gz$", - "errorMessage": "Path to assembly contigs in FASTA format compressed with gzip", - "description": "Metagenomic assembly FASTA file" + "errorMessage": "FASTA file must be provided and have extension '.fa.gz', '.fasta.gz', '.fna.gz'", + "description": "Path to assembly contigs in FASTA format compressed with gzip" }, "fastq_1": { "anyOf": [ diff --git a/assets/schema_input_genome.json b/assets/schema_input_genome.json index 2187d86..4a01222 100644 --- a/assets/schema_input_genome.json +++ b/assets/schema_input_genome.json @@ -19,12 +19,12 @@ "format": "file-path", "exists": true, "pattern": "^([\\S\\s]*\\/)?[^\\s\\/]+\\.(fa|fasta|fna)\\.gz$", - "errorMessage": "Path to MAG/bin sequences in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. File should contain at least one sequence", - "description": "MAG/bin sequence file" + "errorMessage": "Path to the FASTA file must be provided, cannot contain spaces and must have extension '.fa.gz', '.fasta.gz', or '.fna.gz'", + "description": "Path to MAG/bin sequences in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. File should contain at least one sequence" }, "accession": { "type": "string", - "description": "ENA accession of the run or metagenomic assembly used to generate the MAG/bin", + "description": "ENA accession of the run or metagenomic assembly used to generate the MAG/bin. Reads or assembly must already be submitted to ENA", "meta": ["accession"] }, "fastq_1": { @@ -73,7 +73,7 @@ }, "binning_parameters": { "type": "string", - "description": "Arguments used to bin data different from default, e.g. -min_contig_length 1000, otherwise put 'default'", + "description": "Arguments used to bin data different from default, e.g. -min_contig_length 1000, otherwise use 'default'", "meta": ["binning_parameters"] }, "stats_generation_software": { @@ -151,7 +151,7 @@ }, "environmental_medium": { "type": "string", - "description": "Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS", + "description": "Material surrounding and in contact with the sample, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS", "meta": ["environmental_medium"] }, "RNA_presence": { diff --git a/assets/schema_input_reads.json b/assets/schema_input_reads.json index 64931d3..c511c3c 100644 --- a/assets/schema_input_reads.json +++ b/assets/schema_input_reads.json @@ -18,7 +18,7 @@ "type": "string", "pattern": "^\\S+$", "errorMessage": "Sample accession must be provided and cannot contain spaces", - "description": "ENA sample accession (starting with SAMEA) of the sample used to generate raw reads", + "description": "ENA sample accession (starting with SAMEA) of the sample used to generate raw reads. Samples must already be submitted to ENA", "meta": ["sample_accession"] }, "fastq_1": { diff --git a/docs/usage.md b/docs/usage.md index a82cec4..c06657d 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -164,7 +164,7 @@ pacbio_run_001,SAMEA7654321,data/pacbio_reads.fastq.gz,,PACBIO_SMRT,PacBio Seque | Column | Required | Description | | ------------------- | -------- | ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | | `id` | Yes | Unique identifier of this particular data entry. Used as an experiment name. | -| `sample_accession` | Yes | ENA sample accession (starting with SAMEA) of the sample used to generate raw reads. | +| `sample_accession` | Yes | ENA sample accession (starting with SAMEA) of the sample used to generate raw reads. Samples must already be submitted to ENA. | | `fastq_1` | Yes | Path to single-end/paired-end forward reads in FASTQ format (optionally gzipped). | | `fastq_2` | No | Path to reverse reads in FASTQ format for paired-end data (optionally gzipped). Leave empty for single-end reads. | | `platform` | Yes | Sequencing platform. Must be one of the ENA controlled vocabulary values listed in [ENA documentation](https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#metadata-validation). Supported values: `ILLUMINA`, `PACBIO_SMRT`, `OXFORD_NANOPORE`, `ION_TORRENT`, `CAPILLARY`, `DNBSEQ`, `ELEMENT`, `GENAPSYS`, `GENEMIND`, `HELICOS`, `LS454`, `BGISEQ`, `ULTIMA`, `VELA_DIAGNOSTICS`. | @@ -301,12 +301,12 @@ mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default, | --------------------------- | ----------- | -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | | `id` | Yes | A unique identifier for this data entry. Must be globally unique within the input dataset. | | `fasta` | Yes | Path to MAG/bin sequences in FASTA format compressed with `gzip`. All names of the FASTA files must be unique to prevent pipeline errors. File should contain at least one sequence. | -| `accession` | Yes | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. | +| `accession` | Yes | ENA accession of the run or metagenomic assembly used to generate the MAG/bin. Reads or assembly must already be submitted to ENA. | | `fastq_1` | Conditional | Path to single-end/paired-end forward reads in FASTQ format (optionally gzipped) used to generate the source metagenomic assembly. Required if `genome_coverage` is not provided. | | `fastq_2` | No | Path to reverse reads in FASTQ format for paired-end data (optionally gzipped) used to generate the source metagenomic assembly. Leave empty for single-end reads. | | `assembly_software` | Yes | Tool name and version that were used to generate the source metagenomic assembly, e.g. MEGAHIT_v1.0, SPAdes_v4.0.0, metaSPAdes_v3.15.0. | | `binning_software` | Yes | Tool name and version that was used to bin data, e.g. MetaBAT2_v1.0, VAMB_2.0, MaxBin2_v1, CONCOCT_v3.0, SemiBin2_v1, COMEBin_v2.3.4, etc. | -| `binning_parameters` | Yes | Arguments used to bin data different from default, e.g. -min_contig_length 1000, otherwise put 'default'. | +| `binning_parameters` | Yes | Arguments used to bin data different from default, e.g. -min_contig_length 1000, otherwise use 'default'. | | `stats_generation_software` | No | Tool, including version, that was used to calculate completeness and contamination, e.g. CheckM2_v1.0.0. | | `completeness` | No | MAG/bin completeness score: the 0<= ratio <= 100 of observed single-copy marker genes to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined. | | `contamination` | No | MAG/bin contamination score: the 0<= ratio <= 100 of observed single-copy marker genes in ≥2 copies to total single-copy marker genes in chosen marker gene set (%). ENA docs: https://ena-docs.readthedocs.io/en/latest/faq/metagenomes.html#how-is-the-quality-of-a-metagenomic-assembly-defined. | @@ -315,7 +315,7 @@ mag_001,data/mag_001.fasta.gz,SRR24458089,,,SPAdes 3.15.5,MetaBAT2 2.15,default, | `co-assembly` | Yes | Whether a co-assembly strategy was used for the initial metagenomic assembly generation. Options: true or false. | | `broad_environment` | Yes | Broad ecological context of the sample, for example 'marine biome', 'desert biome'. It is recommended to use subclasses of EnvO 'biome' class (http://purl.obolibrary.org/obo/ENVO_00000428). Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS. | | `local_environment` | Yes | Local environmental context of the sample, for example 'tropical dry broadleaf forest biome', 'marine abyssal zone biome'. It is recommended to use EnvO terms which are of smaller spatial grain than your entry for "broad-scale environmental context". Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS. | -| `environmental_medium` | Yes | Material displaced by the sample, or the material in which the sample was embedded before sampling, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS. | +| `environmental_medium` | Yes | Material surrounding and in contact with the sample, for example 'mucus', 'lake water'. It is recommended to use subclasses of EnvO 'environmental material' class (http://purl.obolibrary.org/obo/ENVO_00010483). Documentation: https://github.com/EnvironmentOntology/envo/wiki/Using-ENVO-with-MIxS. | | `RNA_presence` | No | Presence or absence of the 23S, 16S, and 5S rRNA genes and at least 18 tRNAs. This is used for MISAG/MIMAG assembly quality classification. Options: true or false. | | `NCBI_lineage` | No | Full NCBI lineage - format: x;y;z. For example, the lineage for E. coli can be: 'Bacteria;Pseudomonadati;Pseudomonadota;Gammaproteobacteria;Enterobacterales;Enterobacteriaceae;Escherichia' or '2;1224;1236;91347;543;561;562'. For more info check https://www.ncbi.nlm.nih.gov/datasets/docs/v2/data-processing/taxonomy-processing/taxonomy/. | diff --git a/nextflow_schema.json b/nextflow_schema.json index 70dce8f..bbd9f5d 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -19,7 +19,7 @@ "mimetype": "text/csv", "pattern": "^\\S+\\.(csv|tsv|yaml|yml|json)$", "description": "Path to samplesheet describing the data to be submitted (supported formats: csv, tsv, yaml, yml, json). Columns/fields depend on the pipeline mode", - "help": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a file in supported format with a set of columns depending on the type of data being submitted. See [usage docs](https://nf-co.re/seqsubmit/usage#samplesheet-input).", + "help_text": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a file in supported format with a set of columns depending on the type of data being submitted. See [usage docs](https://nf-co.re/seqsubmit/usage#samplesheet-input).", "fa_icon": "fas fa-file-csv" }, "outdir": { @@ -32,7 +32,7 @@ "type": "string", "description": "Email address for completion summary.", "fa_icon": "fas fa-envelope", - "help": "Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (`~/.nextflow/config`) then you don't need to specify this on the command line for every run.", + "help_text": "Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (`~/.nextflow/config`) then you don't need to specify this on the command line for every run.", "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$" }, "multiqc_title": { @@ -47,7 +47,7 @@ "type": "object", "fa_icon": "fas fa-university", "description": "Parameters affecting a decision about presence/absence of tRNA and rRNA genes (RNA_presence) in a genome", - "help": "The value of RNA_presence is based on the number of rRNA and tRNA. If at least 18 tRNAs are detected and 16S, 23S, and 5S rRNA subunits pass the recovery threshold - RNA_presence is True, otherwise - False. The resulting value is a decisive factor affecting MISAG/MIMAG assembly quality classification of a genome.", + "help_text": "The value of RNA_presence is based on the number of rRNA and tRNA. If at least 18 tRNAs are detected and 16S, 23S, and 5S rRNA subunits pass the recovery threshold - RNA_presence is True, otherwise - False. The resulting value is a decisive factor affecting MISAG/MIMAG assembly quality classification of a genome.", "properties": { "rrna_limit": { "type": "number", @@ -68,12 +68,12 @@ "type": "object", "fa_icon": "fas fa-university", "description": "CAT_pack required data", - "help": "Provide a path to pre-downloaded DB or DB id for automatic download", + "help_text": "Provide a path to pre-downloaded DB or DB id for automatic download", "properties": { "cat_db": { "format": "file-path", "description": "Path to local CAT_pack DB", - "help": "Database should be pre-downloaded using CATpack commands. The folder with database should contain sub-folders 'db' with files .dmnd, .fastaid2LCAtaxid, .taxids_with_multiple_offspring, etc, as well as subfolder 'tax' that should contain taxonomy files like .dmp, .taxdump.tar.gz, .prot.accession2taxid.FULL.gz, etc", + "help_text": "Database should be pre-downloaded using CATpack commands. The folder with database should contain sub-folders 'db' with files .dmnd, .fastaid2LCAtaxid, .taxids_with_multiple_offspring, etc, as well as subfolder 'tax' that should contain taxonomy files like .dmp, .taxdump.tar.gz, .prot.accession2taxid.FULL.gz, etc", "fa_icon": "fas fa-users-cog" } } @@ -83,7 +83,7 @@ "type": "object", "fa_icon": "fas fa-university", "description": "Parameters used to run Checkm2", - "help": "These options are used to run Checkm2 that will calculate completeness and contamination of genome if those were not provided as input", + "help_text": "These options are used to run Checkm2 that will calculate completeness and contamination of genome if those were not provided as input", "properties": { "checkm2_db": { "format": "file-path", @@ -103,7 +103,7 @@ "type": "object", "fa_icon": "fas fa-university", "description": "Parameters used to describe centralised config profiles. These should not be edited.", - "help": "The centralised nf-core configuration profiles use a handful of pipeline parameters to describe themselves. This information is then printed to the Nextflow log when you run a pipeline. You should not need to change these values when you run a pipeline.", + "help_text": "The centralised nf-core configuration profiles use a handful of pipeline parameters to describe themselves. This information is then printed to the Nextflow log when you run a pipeline. You should not need to change these values when you run a pipeline.", "properties": { "custom_config_version": { "type": "string", @@ -117,7 +117,7 @@ "description": "Base directory for Institutional configs.", "default": "https://raw.githubusercontent.com/nf-core/configs/master", "hidden": true, - "help": "If you're running offline, Nextflow will not be able to fetch the institutional config files from the internet. If you don't need them, then this is not a problem. If you do need them, you should download the files from the repo and tell Nextflow where to find them with this parameter.", + "help_text": "If you're running offline, Nextflow will not be able to fetch the institutional config files from the internet. If you don't need them, then this is not a problem. If you do need them, you should download the files from the repo and tell Nextflow where to find them with this parameter.", "fa_icon": "fas fa-users-cog" }, "config_profile_name": { @@ -151,7 +151,7 @@ "type": "object", "fa_icon": "fas fa-file-import", "description": "Less common options for the pipeline, typically set in a config file.", - "help": "These options are common to all nf-core pipelines and allow you to customise some of the core preferences for how the pipeline runs.\n\nTypically these options would be set in a Nextflow config file loaded for all pipeline runs, such as `~/.nextflow/config`.", + "help_text": "These options are common to all nf-core pipelines and allow you to customise some of the core preferences for how the pipeline runs.\n\nTypically these options would be set in a Nextflow config file loaded for all pipeline runs, such as `~/.nextflow/config`.", "properties": { "version": { "type": "boolean", @@ -163,7 +163,7 @@ "type": "string", "default": "copy", "description": "Method used to save pipeline results to output directory.", - "help": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", + "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", "fa_icon": "fas fa-copy", "enum": ["symlink", "rellink", "link", "copy", "copyNoFollow", "move"], "hidden": true @@ -173,7 +173,7 @@ "description": "Email address for completion summary, only when pipeline fails.", "fa_icon": "fas fa-exclamation-triangle", "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$", - "help": "An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully.", + "help_text": "An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully.", "hidden": true }, "plaintext_email": { @@ -257,50 +257,50 @@ "centre_name": { "type": "string", "description": "Name of the submitter's organisation (mandatory for broker accounts).", - "help": "ena-webin-cli argument -centerName: the center name of the submitter (mandatory for broker accounts)." + "help_text": "ena-webin-cli argument -centerName: the center name of the submitter (mandatory for broker accounts)." }, "upload_tpa": { "type": "boolean", "description": "Mark the assemblies as a Third PArty (TPA)", - "help": "Set to true if the assembly was built from public sequence data (TPA) rather than original primary data. See NCBI TPA guidance: https://www.ncbi.nlm.nih.gov/genbank/tpa/", + "help_text": "Set to true if the assembly was built from public sequence data (TPA) rather than original primary data. See NCBI TPA guidance: https://www.ncbi.nlm.nih.gov/genbank/tpa/", "default": false }, "mode": { "type": "string", "default": null, "description": "Type of the data to be submitted", - "help": "Different types of data require specific upload steps. That mode controls what upload workflow to run depending on type of data (mags/bins/metagenomic_assemblies/reads)", + "help_text": "Different types of data require specific upload steps. That mode controls what upload workflow to run depending on type of data (mags/bins/metagenomic_assemblies/reads)", "enum": ["mags", "bins", "metagenomic_assemblies", "reads"] }, "test_upload": { "type": "boolean", "description": "Upload to ENA TEST server instead of LIVE server", "default": true, - "help": "Use that flag for development purposes or for validation of your submissions. Submission would be done to TEST ENA server instead of LIVE server. TEST server holds data 24h and removes afterwords. LIVE server does not have option to modify your submission, you can only suppress data and re-upload" + "help_text": "Use that flag for development purposes or for validation of your submissions. Submission would be done to TEST ENA server instead of LIVE server. TEST server holds data 24h and removes afterwords. LIVE server does not have option to modify your submission, you can only suppress data and re-upload" }, "is_private": { "type": "boolean", "default": false, "description": "Use that flag if you are referencing private data accessions in the samplesheet", - "help": "Set to true when the referenced project/reads/assembly are still private in ENA, so metadata is fetched via authenticated access. Does NOT affect the privacy of your own submission, use --release_date for that" + "help_text": "Set to true when the referenced project/reads/assembly are still private in ENA, so metadata is fetched via authenticated access. Does NOT affect the privacy of your own submission, use --release_date for that" }, "release_date": { "type": "string", "pattern": "^\\d{4}-\\d{2}-\\d{2}$", "description": "Date (YYYY-MM-DD) until which the newly created study stays private in ENA.", - "help": "Sets the release date for the study created by the pipeline. Data stays private until this date, then is automatically released and indexed by ENA. Must be no more than 2 years from today. Omit to release the study publicly immediately." + "help_text": "Sets the release date for the study created by the pipeline. Data stays private until this date, then is automatically released and indexed by ENA. Must be no more than 2 years from today. Omit to release the study publicly immediately." }, "submission_study": { "type": "string", "description": "ENA study accession (PRJ/ERP) to submit the data to", - "help": "Current implementation of pipeline requires to pre-register ENA project (PRJ/ERP) where you want to upload data to. Documentation how to register study: https://ena-docs.readthedocs.io/en/latest/submit/study.html" + "help_text": "Current implementation of pipeline requires to pre-register ENA project (PRJ/ERP) where you want to upload data to. Documentation how to register study: https://ena-docs.readthedocs.io/en/latest/submit/study.html" }, "study_metadata": { "type": "string", "format": "file-path", "exists": true, "description": "Path to a file (JSON, CSV, or TSV) with metadata for registering a new ENA study. Required if --submission_study is not set.", - "help": "File containing study metadata fields (required: study_title, alias; optional: study_abstract, existing_study_type, etc.) used by the pipeline to register a new ENA study when no existing --submission_study accession is given. ENA docs: https://ena-docs.readthedocs.io/en/latest/submit/study/programmatic.html", + "help_text": "File containing study metadata fields (required: study_title, alias; optional: study_abstract, existing_study_type, etc.) used by the pipeline to register a new ENA study when no existing --submission_study accession is given. ENA docs: https://ena-docs.readthedocs.io/en/latest/submit/study/programmatic.html", "fa_icon": "fas fa-file-alt" }, "webincli_mode": { @@ -308,7 +308,7 @@ "description": "Webin-CLI mode for ENA interaction: `submit` uploads data, `validate` performs validation only.", "default": "submit", "enum": ["submit", "validate"], - "help": "Controls ENA Webin-CLI mode. `validate` mode checks correctness of manifest files, data access and validity. `submit` mode does validation and push data to ENA server. Default: submit." + "help_text": "Controls ENA Webin-CLI mode. `validate` mode checks correctness of manifest files, data access and validity. `submit` mode does validation and push data to ENA server. Default: submit." } } } diff --git a/ro-crate-metadata.json b/ro-crate-metadata.json index 7b73754..5bf02a8 100644 --- a/ro-crate-metadata.json +++ b/ro-crate-metadata.json @@ -23,7 +23,7 @@ "@type": "Dataset", "creativeWorkStatus": "InProgress", "datePublished": "2026-04-30T13:32:49+00:00", - "description": "

    \n \n \n \"nf-core/seqsubmit\"\n \n

    \n\n[![Open in GitHub Codespaces](https://img.shields.io/badge/Open_In_GitHub_Codespaces-black?labelColor=grey&logo=github)](https://github.com/codespaces/new/nf-core/seqsubmit)\n[![GitHub Actions CI Status](https://github.com/nf-core/seqsubmit/actions/workflows/nf-test.yml/badge.svg)](https://github.com/nf-core/seqsubmit/actions/workflows/nf-test.yml)\n[![GitHub Actions Linting Status](https://github.com/nf-core/seqsubmit/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/seqsubmit/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/seqsubmit/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX)\n[![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com)\n\n[![Nextflow](https://img.shields.io/badge/version-%E2%89%A525.10.4-green?style=flat&logo=nextflow&logoColor=white&color=%230DC09D&link=https%3A%2F%2Fnextflow.io)](https://www.nextflow.io/)\n[![nf-core template version](https://img.shields.io/badge/nf--core_template-4.0.2-green?style=flat&logo=nfcore&logoColor=white&color=%2324B064&link=https%3A%2F%2Fnf-co.re)](https://github.com/nf-core/tools/releases/tag/4.0.2)\n[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/)\n[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/)\n[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/)\n[![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/seqsubmit)\n\n[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23seqsubmit-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/seqsubmit)[![Follow on Bluesky](https://img.shields.io/badge/bluesky-%40nf__core-1185fe?labelColor=000000&logo=bluesky)](https://bsky.app/profile/nf-co.re)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core)\n\n## Introduction\n\n**nf-core/seqsubmit** is a Nextflow pipeline for submitting sequence data to [ENA](https://www.ebi.ac.uk/ena/browser/home).\nCurrently, the pipeline supports four submission modes, each routed to a dedicated workflow and requiring its own input samplesheet structure:\n\n- `mags` for Metagenome Assembled Genomes (MAGs) submission with `GENOMESUBMIT` workflow\n- `bins` for bins submission with `GENOMESUBMIT` workflow\n- `metagenomic_assemblies` for assembly submission with `ASSEMBLYSUBMIT` workflow\n- `reads` for raw sequencing reads submission with `READSUBMIT` workflow\n\n![seqsubmit workflow diagram](docs/images/seqsubmit_schema.png)\n\n**nf-core/seqsubmit** is a Nextflow pipeline for submitting sequence data to [ENA](https://www.ebi.ac.uk/ena/browser/home).\nThe pipeline currently supports the following submission modes, each routed to a dedicated workflow:\n\n- `reads` \u2014 raw sequencing reads submission via the `READSUBMIT` workflow (pink)\n- `metagenomic_assemblies` \u2014 assembly submission via the `ASSEMBLYSUBMIT` workflow (green)\n- `mags` \u2014 metagenome-assembled genomes (MAGs) submission via the `GENOMESUBMIT` workflow (blue)\n- `bins` \u2014 bins submission via the `GENOMESUBMIT` workflow (blue)\n\n\n\nEach workflow has its own samplesheet structure, prerequisites, and limitations \u2014 they are briefly described below. See the [usage documentation](https://nf-co.re/seqsubmit/usage) for more detailed explanations.\n\n## Requirements\n\n- [Nextflow](https://www.nextflow.io/) `>=25.04.0`\n- A Webin account registered at \n\n Set your Webin credentials as Nextflow secrets:\n\n ```bash\n nextflow secrets set ENA_WEBIN \"Webin-XXX\"\n nextflow secrets set ENA_WEBIN_PASSWORD \"XXX\"\n ```\n\n Make sure to replace the values above with your own credentials.\n\n- Provide either a study accession or a study registration metadata file for the study the submission will be associated with. See the [Submission study](docs/usage.md#submission-study) section of the usage documentation for details.\n\n- Depending on the chosen mode, samples, reads, or metagenomic assemblies must be pre-submitted to ENA to obtain the corresponding accessions, which you then reference in your submission. Refer to the relevant mode section in the [usage documentation](https://nf-co.re/seqsubmit/usage) for details.\n\n## Input samplesheets\n\n### `reads` mode\n\nExample:\n\n```csv\nsample,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,library_selection,library_strategy,insert_size,library_name,description\nillumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLUMINA,Illumina HiSeq 2000,GENOMIC,RANDOM,WGS,500,HiSeq_library_001,Illumina sequencing of sample XYZ\n```\n\nSee the [`reads` mode section](docs/usage.md#samplesheet-input) of the usage documentation for more details.\n\n### `metagenomic_assemblies` mode\n\nExample:\n\n```csv\nsample,fasta,fastq_1,fastq_2,coverage,run_accession,assembler,assembler_version\nassembly_1,data/contigs_1.fasta.gz,data/reads_1.fastq.gz,data/reads_2.fastq.gz,,ERR011322,SPAdes,3.15.5\nassembly_2,data/contigs_2.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9\n```\n\nSee the [`metagenomic_assemblies` mode section](docs/usage.md#samplesheet-input-1) of the usage documentation for more details.\n\n### `mags` and `bins` modes\n\nExample:\n\n```csv\nsample,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage\nlachnospira_eligens,data/bin_lachnospira_eligens.fa.gz,SRR24458089,,,spades_v3.15.5,metabat2_v2.6,default,CheckM2_v1.0.1,61.0,0.21,32.07,sediment metagenome,No,marine,cable_bacteria,marine_sediment,No,d__Bacteria;p__Proteobacteria;s__unclassified_Proteobacteria\n```\n\nSee the [`mags` and `bins` modes section](docs/usage.md#samplesheet-input-2) of the usage documentation for the full list of required and optional columns.\n\n## Usage\n\n> [!NOTE]\n> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/get_started/environment_setup/overview) on how to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/get_started/run-your-first-pipeline) with `-profile test` before running the workflow on actual data.\n\n### Running the pipeline\n\nEach mode also has its own additional parameters and example commands \u2014 see the [usage documentation](https://nf-co.re/seqsubmit/usage) for details. General command template:\n\n```bash\nnextflow run nf-core/seqsubmit \\\n -profile \\\n --mode \\\n --input \\\n --centre_name \\\n --submission_study \\\n --outdir \n```\n\n> [!WARNING]\n> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; see [docs](https://nf-co.re/docs/running/run-pipelines#using-parameter-files).\n\nFor more details and further functionality, please refer to the [usage documentation](https://nf-co.re/seqsubmit/usage) and the [parameter documentation](https://nf-co.re/seqsubmit/parameters).\n\n## Pipeline output\n\nKey output locations in `--outdir`:\n\n- `reads/`: per-sample submission receipts and accessions\n- `metagenomic_assemblies/`: assembly metadata CSVs and per-sample coverage files\n- `mags/` or `bins/`: genome metadata, manifests, and per-sample submission support files\n- `multiqc/`: MultiQC summary report\n- `pipeline_info/`: execution reports, trace, DAG, and software versions\n\nFor full details, see the [output documentation](https://nf-co.re/seqsubmit/output).\n\n## Credits\n\nnf-core/seqsubmit was originally written by [Martin Beracochea](https://github.com/mberacochea), [Ekaterina Sakharova](https://github.com/KateSakharova), [Sofia Ochkalova](https://github.com/ochkalova), [Evangelos Karatzas](https://github.com/vagkaratzas) and [Tim Rozday](https://github.com/timrozday-mgnify).\n\n## Contributions and Support\n\nIf you would like to contribute to this pipeline, please see the [contributing guidelines](docs/CONTRIBUTING.md).\n\nFor further information or help, don't hesitate to get in touch on the [Slack `#seqsubmit` channel](https://nfcore.slack.com/channels/seqsubmit) (you can join with [this invite](https://nf-co.re/join/slack)).\n\n## Citations\n\n\n\n\n\nIf you use this pipeline please make sure to cite all used software.\nThis pipeline uses code and infrastructure developed and maintained by the [nf-core](https://nf-co.re) community, reused here under the [MIT license](https://github.com/nf-core/tools/blob/main/LICENSE).\n\n> **MGnify: the microbiome sequence data analysis resource in 2023**\n>\n> Richardson L, Allen B, Baldi G, Beracochea M, Bileschi ML, Burdett T, et al.\n>\n> Vol. 51, Nucleic Acids Research. Oxford University Press (OUP); 2022. p. D753\u20139. Available from: http://dx.doi.org/10.1093/nar/gkac1080\n\nAn extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file.\n\nYou can cite the `nf-core` publication as follows:\n\n> **The nf-core framework for community-curated bioinformatics pipelines.**\n>\n> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen.\n>\n> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x).\n", + "description": "

    \n \n \n \"nf-core/seqsubmit\"\n \n

    \n\n[![Open in GitHub Codespaces](https://img.shields.io/badge/Open_In_GitHub_Codespaces-black?labelColor=grey&logo=github)](https://github.com/codespaces/new/nf-core/seqsubmit)\n[![GitHub Actions CI Status](https://github.com/nf-core/seqsubmit/actions/workflows/nf-test.yml/badge.svg)](https://github.com/nf-core/seqsubmit/actions/workflows/nf-test.yml)\n[![GitHub Actions Linting Status](https://github.com/nf-core/seqsubmit/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/seqsubmit/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/seqsubmit/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX)\n[![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com)\n\n[![Nextflow](https://img.shields.io/badge/version-%E2%89%A525.10.4-green?style=flat&logo=nextflow&logoColor=white&color=%230DC09D&link=https%3A%2F%2Fnextflow.io)](https://www.nextflow.io/)\n[![nf-core template version](https://img.shields.io/badge/nf--core_template-4.0.2-green?style=flat&logo=nfcore&logoColor=white&color=%2324B064&link=https%3A%2F%2Fnf-co.re)](https://github.com/nf-core/tools/releases/tag/4.0.2)\n[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/)\n[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/)\n[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/)\n[![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/seqsubmit)\n\n[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23seqsubmit-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/seqsubmit)[![Follow on Bluesky](https://img.shields.io/badge/bluesky-%40nf__core-1185fe?labelColor=000000&logo=bluesky)](https://bsky.app/profile/nf-co.re)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core)\n\n## Introduction\n\n![seqsubmit workflow diagram](docs/images/seqsubmit_schema.png)\n\n**nf-core/seqsubmit** is a Nextflow pipeline for submitting sequence data to [ENA](https://www.ebi.ac.uk/ena/browser/home).\nThe pipeline currently supports the following submission modes, each routed to a dedicated workflow:\n\n- `reads` \u2014 raw sequencing reads submission via the `READSUBMIT` workflow (pink)\n- `metagenomic_assemblies` \u2014 assembly submission via the `ASSEMBLYSUBMIT` workflow (green)\n- `mags` \u2014 metagenome-assembled genomes (MAGs) submission via the `GENOMESUBMIT` workflow (blue)\n- `bins` \u2014 bins submission via the `GENOMESUBMIT` workflow (blue)\n\n\n\nEach workflow has its own samplesheet structure, prerequisites, and limitations \u2014 they are briefly described below. See the [usage documentation](https://nf-co.re/seqsubmit/usage) for more detailed explanations.\n\n## Requirements\n\n- [Nextflow](https://www.nextflow.io/) `>=25.04.0`\n- A Webin account registered at \n\n Set your Webin credentials as Nextflow secrets:\n\n ```bash\n nextflow secrets set ENA_WEBIN \"Webin-XXX\"\n nextflow secrets set ENA_WEBIN_PASSWORD \"XXX\"\n ```\n\n Make sure to replace the values above with your own credentials.\n\n- Provide either a study accession or a study registration metadata file for the study the submission will be associated with. See the [Submission study](docs/usage.md#submission-study) section of the usage documentation for details.\n\n- Depending on the chosen mode, samples, reads, or metagenomic assemblies must be pre-submitted to ENA to obtain the corresponding accessions, which you then reference in your submission. Refer to the relevant mode section in the [usage documentation](https://nf-co.re/seqsubmit/usage) for details.\n\n## Input samplesheets\n\n### `reads` mode\n\nExample:\n\n```csv\nid,sample_accession,fastq_1,fastq_2,platform,instrument,library_source,library_selection,library_strategy,insert_size,library_name,description\nillumina_run_001,SAMEA1234567,data/reads_R1.fastq.gz,data/reads_R2.fastq.gz,ILLUMINA,Illumina HiSeq 2000,GENOMIC,RANDOM,WGS,500,HiSeq_library_001,Illumina sequencing of sample XYZ\n```\n\nSee the [`reads` mode section](docs/usage.md#samplesheet-input) of the usage documentation for more details.\n\n### `metagenomic_assemblies` mode\n\nExample:\n\n```csv\nid,fasta,fastq_1,fastq_2,coverage,run_accession,assembler,assembler_version\nassembly_1,data/contigs_1.fasta.gz,data/reads_1.fastq.gz,data/reads_2.fastq.gz,,ERR011322,SPAdes,3.15.5\nassembly_2,data/contigs_2.fasta.gz,,,42.7,ERR011323,MEGAHIT,1.2.9\n```\n\nSee the [`metagenomic_assemblies` mode section](docs/usage.md#samplesheet-input-1) of the usage documentation for more details.\n\n### `mags` and `bins` modes\n\nExample:\n\n```csv\nid,fasta,accession,fastq_1,fastq_2,assembly_software,binning_software,binning_parameters,stats_generation_software,completeness,contamination,genome_coverage,metagenome,co-assembly,broad_environment,local_environment,environmental_medium,RNA_presence,NCBI_lineage\nlachnospira_eligens,data/bin_lachnospira_eligens.fa.gz,SRR24458089,,,spades_v3.15.5,metabat2_v2.6,default,CheckM2_v1.0.1,61.0,0.21,32.07,sediment metagenome,false,marine,cable_bacteria,marine_sediment,false,d__Bacteria;p__Proteobacteria;s__unclassified_Proteobacteria\n```\n\nSee the [`mags` and `bins` modes section](docs/usage.md#samplesheet-input-2) of the usage documentation for the full list of required and optional columns.\n\n## Usage\n\n> [!NOTE]\n> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/get_started/environment_setup/overview) on how to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/get_started/run-your-first-pipeline) with `-profile test` before running the workflow on actual data.\n\n### Running the pipeline\n\nEach mode also has its own additional parameters and example commands \u2014 see the [usage documentation](https://nf-co.re/seqsubmit/usage) for details. General command template:\n\n```bash\nnextflow run nf-core/seqsubmit \\\n -profile \\\n --mode \\\n --input \\\n --centre_name \\\n --submission_study \\\n --outdir \n```\n\n> [!WARNING]\n> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; see [docs](https://nf-co.re/docs/running/run-pipelines#using-parameter-files).\n\nFor more details and further functionality, please refer to the [usage documentation](https://nf-co.re/seqsubmit/usage) and the [parameter documentation](https://nf-co.re/seqsubmit/parameters).\n\n## Pipeline output\n\nKey output locations in `--outdir`:\n\n- `reads/`: per-sample submission receipts and accessions\n- `metagenomic_assemblies/`: assembly metadata CSVs and per-sample coverage files\n- `mags/` or `bins/`: genome metadata, manifests, and per-sample submission support files\n- `multiqc/`: MultiQC summary report\n- `pipeline_info/`: execution reports, trace, DAG, and software versions\n\nFor full details, see the [output documentation](https://nf-co.re/seqsubmit/output).\n\n## Credits\n\nnf-core/seqsubmit was originally written by [Martin Beracochea](https://github.com/mberacochea), [Ekaterina Sakharova](https://github.com/KateSakharova), [Sofia Ochkalova](https://github.com/ochkalova), [Evangelos Karatzas](https://github.com/vagkaratzas) and [Tim Rozday](https://github.com/timrozday-mgnify).\n\n## Contributions and Support\n\nIf you would like to contribute to this pipeline, please see the [contributing guidelines](docs/CONTRIBUTING.md).\n\nFor further information or help, don't hesitate to get in touch on the [Slack `#seqsubmit` channel](https://nfcore.slack.com/channels/seqsubmit) (you can join with [this invite](https://nf-co.re/join/slack)).\n\n## Citations\n\n\n\n\n\nIf you use this pipeline please make sure to cite all used software.\nThis pipeline uses code and infrastructure developed and maintained by the [nf-core](https://nf-co.re) community, reused here under the [MIT license](https://github.com/nf-core/tools/blob/main/LICENSE).\n\n> **MGnify: the microbiome sequence data analysis resource in 2023**\n>\n> Richardson L, Allen B, Baldi G, Beracochea M, Bileschi ML, Burdett T, et al.\n>\n> Vol. 51, Nucleic Acids Research. Oxford University Press (OUP); 2022. p. D753\u20139. Available from: http://dx.doi.org/10.1093/nar/gkac1080\n\nAn extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file.\n\nYou can cite the `nf-core` publication as follows:\n\n> **The nf-core framework for community-curated bioinformatics pipelines.**\n>\n> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen.\n>\n> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x).\n", "hasPart": [ { "@id": "main.nf" From 19067fb21965caddafd1954a4998de77edb53d4e Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Thu, 30 Jul 2026 14:38:15 +0100 Subject: [PATCH 45/47] remove files_unchanged --- .nf-core.yml | 5 ----- 1 file changed, 5 deletions(-) diff --git a/.nf-core.yml b/.nf-core.yml index bf38907..fc8d70b 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -5,11 +5,6 @@ lint: - conf/igenomes_ignored.config nextflow_config: - params.input - files_unchanged: - - .github/PULL_REQUEST_TEMPLATE.md - - docs/images/nf-core-seqsubmit_logo_light.png - - docs/images/nf-core-seqsubmit_logo_dark.png - - assets/nf-core-seqsubmit_logo_light.png nf_core_version: 4.0.3 repository_type: pipeline template: From decd115d75ae4d17dbffd5a11772fac05a84a03b Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova Date: Thu, 30 Jul 2026 14:44:55 +0100 Subject: [PATCH 46/47] revert files_unchanged --- .nf-core.yml | 5 +++++ CHANGELOG.md | 2 +- 2 files changed, 6 insertions(+), 1 deletion(-) diff --git a/.nf-core.yml b/.nf-core.yml index fc8d70b..bf38907 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -5,6 +5,11 @@ lint: - conf/igenomes_ignored.config nextflow_config: - params.input + files_unchanged: + - .github/PULL_REQUEST_TEMPLATE.md + - docs/images/nf-core-seqsubmit_logo_light.png + - docs/images/nf-core-seqsubmit_logo_dark.png + - assets/nf-core-seqsubmit_logo_light.png nf_core_version: 4.0.3 repository_type: pipeline template: diff --git a/CHANGELOG.md b/CHANGELOG.md index d12e871..a716c4a 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). -## v1.0.0 - [date] +## v1.0.0 - 01/08/2026 Initial release of nf-core/seqsubmit, created with the [nf-core](https://nf-co.re/) template. From 670bdc646eaa09ae6f19361423a7f735705528a8 Mon Sep 17 00:00:00 2001 From: Ekaterina Sakharova <49755902+KateSakharova@users.noreply.github.com> Date: Thu, 30 Jul 2026 15:28:03 +0100 Subject: [PATCH 47/47] Revise resource settings guidance in usage documentation Updated resource settings reference for Nextflow configuration. --- docs/usage.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/usage.md b/docs/usage.md index c06657d..14c38e2 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -386,7 +386,7 @@ nextflow run nf-core/seqsubmit \ Some processes in this pipeline are resource intensive — in particular coverage calculation (CoverM), quality assessment (CheckM2), and taxonomic classification of MAGs/bins (BAT). For real-world data, these steps should be run on an HPC cluster or another compute environment with sufficient CPU, memory, and job scheduling capacity, rather than on a single local machine. -Depending on the resources available on your machine or cluster, you will likely need to adapt the Nextflow configuration to limit the maximum number of concurrently running jobs and other resource settings (e.g. `max_cpus`, `max_memory`, `max_time`, or `executor.queueSize` in a custom config), so that the pipeline does not try to claim more resources than are actually available. See the [nf-core configuration documentation](https://nf-co.re/docs/usage/getting_started/configuration) for guidance on writing a custom config for your infrastructure. +Depending on the resources available on your machine or cluster, you will likely need to adapt the Nextflow configuration to limit the maximum number of concurrently running jobs and other resource settings using [resourceLimits](https://nf-co.re/docs/developing/institutional-profiles/configuration#resource-limits) in a custom config, so that the pipeline does not try to claim more resources than are actually available. See the [nf-core configuration documentation](https://nf-co.re/docs/usage/getting_started/configuration) for guidance on writing a custom config for your infrastructure. When submitting a large number of records, be aware that ENA recommends limiting submissions to 5000 per day to avoid overfilling their processing queue. If you need to submit more than this, consider splitting your samplesheet and spreading the submission across several days.