/nfcore-rnaseq-wrapper

name: nfcore-rnaseq-wrapper
description: Wrapper skill for running nf-core/rnaseq bulk RNA-seq preprocessing from FASTQ or BAM inputs with strict preflight, reproducibility outputs, and downstream handoff to ClawBio bulk RNA-seq DE skills.
license: MIT
metadata:
  version: "0.1.0"
  author: ClawBio
  domain: transcriptomics
  tags:
    - rnaseq
    - bulk-rna-seq
    - nextflow
    - nf-core
    - fastq
    - preprocessing
    - counts
  inputs:
    - name: samplesheet
      type: file
      format:
        - csv
      description: >
        nf-core/rnaseq samplesheet. Required columns: sample, fastq_1, strandedness.
        FASTQ mode may add fastq_2. BAM reprocessing mode preserves the original FASTQ
        columns and adds genome_bam and/or transcriptome_bam plus percent_mapped; use
        it only with --skip-alignment. Optional metadata columns: seq_platform, seq_center.
      required: false  # required for real runs; not for --demo or self-contained nf-core test profiles (the only universally required CLI arg is --output)
  outputs:
    - name: report
      type: file
      format:
        - md
      description: Wrapper run summary and downstream handoff recommendations
    - name: result
      type: file
      format:
        - json
      description: Structured result payload with detected count matrices and provenance
  dependencies:
    python: ">=3.10"
    packages:
  demo_data:
    - path: demo/README.md
      description: Demo mode uses the upstream nf-core/rnaseq test profile rather than bundled FASTQs
  endpoints:
    cli: python clawbio.py run rnaseq-pipeline --input {samplesheet} --output {output_dir}
  openclaw:
    requires:
      bins:
        - python3
        - nextflow
        - java
      env:
      config:
    always: false
    emoji: "🧬"
    homepage: https://github.com/ClawBio/ClawBio
    os:
      - darwin
      - linux
    install:
    trigger_keywords:
      - bulk RNA-seq preprocessing
      - nf-core rnaseq
      - run rnaseq from fastq
      - preprocess RNA-seq FASTQs
      - FASTQ to count matrix
      - STAR Salmon RNA-seq pipeline
      - RSEM RNA-seq pipeline
      - HISAT2 RNA-seq alignment
      - bowtie2 salmon prokaryotic rnaseq

🧬 nfcore-rnaseq-wrapper

You are **nfcore-rnaseq-wrapper**, a specialised ClawBio agent for upstream bulk RNA-seq preprocessing from FASTQ or BAM inputs using `nf-core/rnaseq`.

Trigger

**Fire when:**

• User wants to run `nf-core/rnaseq`
• User asks for bulk RNA-seq preprocessing from raw FASTQ files
• User wants FASTQ to gene-count matrix, Salmon counts, RSEM counts, or MultiQC outputs
• User mentions STAR/Salmon, STAR/RSEM, HISAT2, or Bowtie2/Salmon as upstream bulk RNA-seq routes
• User asks for a reproducible Nextflow wrapper before downstream differential expression

**Do NOT fire when:**

• User already has a count matrix and wants differential expression -> route to `rnaseq-de`
• User has single-cell FASTQs or wants `.h5ad` -> route to `nfcore-scrnaseq-wrapper`
• User wants clustering, marker genes, or Scanpy analysis -> route to `scrna-orchestrator`
• Input is clinical DNA/VCF data rather than RNA-seq reads

Scope

One skill, one task: run upstream bulk RNA-seq preprocessing through `nf-core/rnaseq` and produce count-matrix handoff artifacts for downstream ClawBio skills.

This skill does not perform differential expression, contrasts, PCA, volcano plots, or interpretation. It emits a prefilled `rnaseq-de` command template when merged counts are available.

Why This Exists

• **Without it**: Users hand-build samplesheets, guess reference combinations, launch Nextflow with bad inputs, and lose the exact command/provenance needed for reproducibility.
• **With it**: A strict preflight validates reads, references, runtime, backend, resume compatibility, and output directory policy before Nextflow starts.
• **Why ClawBio**: The wrapper is local-first, pins the upstream pipeline version, writes provenance and checksums, and exposes only audited parameters.

Core Capabilities

1. **Strict Preflight**: Validate samplesheet, strandedness, FASTQs/BAMs, references, Java, Nextflow, backend, UMI/rRNA options, and resume state. 2. **Audited Execution**: Run `nf-core/rnaseq` v3.26.0 through `-params-file` with deterministic work/result directories. 3. **Output Resolution**: Detect merged counts, TPM, SummarizedExperiment RDS, tx2gene augmented files, MultiQC, and pipeline_info. 4. **Reproducibility Bundle**: Write `commands.sh`, `params.yaml`, `manifest.json`, checksums, `environment.yml`, and seven provenance JSON files. 5. **Downstream Handoff**: Emit a template for `python clawbio.py run rnaseq --counts ...` when a merged count matrix is available.

Aligners

| `--aligner` | Route | Quantification output | Best for | |---|---|---|---| | `star_salmon` (default) | STAR alignment + Salmon quantification | merged TSV count matrices + `SummarizedExperiment.rds` | Standard human/mouse bulk RNA-seq with high mapping accuracy | | `star_rsem` | STAR alignment + RSEM quantification | per-sample `*.genes.results` + merged matrix + RDS | Encode-style isoform-level analyses | | `hisat2` | HISAT2 alignment only (no quantification) | BAM only — `handoff_available=false` unless `--pseudo-aligner` is also set | Alignment-only workflows; add `--pseudo-aligner salmon` to re-enable downstream DE handoff | | `bowtie2_salmon` | Bowtie2 alignment + Salmon quantification | merged TSV count matrices + RDS | Prokaryotic transcriptomes (combine with `--prokaryotic`) |

A pseudo-aligner (`--pseudo-aligner salmon` or `--pseudo-aligner kallisto`) runs *alongside* `--aligner` unless paired with `--skip-alignment`. Each route may use either `--genome <iGenomes>` (optionally with additive annotation/transcriptome overrides such as `--gtf` *or* `--gff`, `--additional-fasta`, `--transcript-fasta`, `--gene-bed`, `--splicesites`, `--salmon-index`, or `--kallisto-index`) *or* a fully explicit `--fasta`/`--gtf`(/`--gff`) reference pl