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fullstack-omics

RESEARCH USE ONLY - not clinically validated.

Full-stack, autonomous single-cell omics: from "run this assay on N cells" all the way to a result. Cheap, fast, autonomous, with molecule counting (UMIs), aimed at expert-level benchmarks on commodity hardware (Hamilton + PyLabRobot + on-deck thermocycler + BioTek Synergy H1).

Each assay is a self-contained module that runs end to end: purchase, instrument bring-up, liquid-handling readiness, automated execution, and fastq-to-analysis. Everything runs in the PyLabRobot simulator with no hardware attached.

Modules

Module Assay Status
autonomous-scRNAseq/ FLASH-seq UMI v3 (low-input full-length scRNA-seq + UMIs) v0.1: 4 stages + Rhodamine B readiness QC + human/humanoid operator + fastq-to-analysis (scanpy), all wired end to end in the PyLabRobot simulator
autonomous-scWGS/ single-cell WGS: ResolveDNA PTA WGA + NEBNext Ultra II (FACS Melody sort up front) v0.1: sort + WGA + library prep + Rhodamine readiness + human/humanoid operator + BJ-WGS sequencing analysis, end to end in the PyLabRobot simulator (10/10 tests)

autonomous-scRNAseq/ is the FLASH-seq scRNA-seq pipeline. It delivers, all runnable with no hardware: procurement (BOM to IDT/browser/PO channels to one human approval), a printable bench manual, instrument-readiness QC (Rhodamine B pipetting precision on a Synergy H1), the full FLASH-seq automation in the simulator with QC gates and tacit guards, a swappable human or humanoid operator, and a fastq-to-analysis pipeline (bcl2fastq to umi_tools to STAR to featureCounts to umi_tools count to scanpy).

autonomous-scWGS/ is the single-cell whole-genome sequencing pipeline: a BD FACS Melody sort into ResolveDNA Cell Buffer, ResolveDNA PTA whole-genome amplification, post-WGA dsDNA QC on the Synergy H1, NEBNext Ultra II library prep (exact SPRI ratios, do-not- over-dry guards), pool, and a fastq-to-variants analysis via the BaseJumper BJ-WGS Nextflow pipeline (Sentieon BWA MEM to Dedup to BQSR to DNAScope to SnpEff/ClinVar/dbSNP to MultiQC). It shares the Rhodamine B readiness QC and human/humanoid operator with the scRNAseq module. The underlying kits are proprietary (RESEARCH USE ONLY), not CC-BY; values are sourced from the vendor guides and never invented. The FACS Melody control plane is being reverse-engineered (owner's TODO); sorting is simulated until wired.

The autonomy stack (shared across modules)

  1. Procurement - resolve reagents to SKUs, route to IDT API / browser carts / PO, one human approval, schedule the run on lead times.
  2. Instrument bring-up - controller kit (Raspberry Pi + PyLabRobot) and a connectivity registry that emits exact cabling. Includes reverse-engineering the cell sorter.
  3. Readiness - Rhodamine B liquid-handling CV check: READY or NEEDS_CALIBRATION.
  4. Execution - the assay on a Hamilton deck + on-deck thermocycler + Synergy H1.
  5. Ops - a human or an (experimental) humanoid robot sets up the deck, presses run, collects output.
  6. Analysis - fastq to count matrix to clustered result (UMI molecule counting).

Known hard subproblem: FACS Melody

The single-cell sort (cells into lysis buffer) depends on a BD FACS Melody, which has no open automation API, so programmatic control has to be reverse-engineered. Tracked per module. Nothing about that interface is invented; unknowns stay TODO.

Principles

  • Never invent a value: source from the protocol; unknowns are # TODO, OCR-ambiguous items are verify, tuning knobs are # expert-tunable.
  • One human approval before any purchase; fail closed on readiness.
  • Swappable backends: everything runs in simulation with no hardware.

License

Code: MIT (LICENSE). Each module credits its source protocol under that protocol's own license (FLASH-seq UMI v3 is CC-BY; see the module's ATTRIBUTION).

About

Full-stack autonomous single-cell omics, end to end in the PyLabRobot simulator. Two modules: autonomous-scRNAseq (FLASH-seq UMI v3) and autonomous-scWGS (ResolveDNA PTA + NEBNext, BJ-WGS analysis). Procurement, Rhodamine readiness QC, automated execution, fastq-to-analysis. RESEARCH USE ONLY.

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