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Copilot AI commented Nov 25, 2025

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
  • If necessary, also make a PR on the nf-core/rnaseq branch on the 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 <OUTDIR>).
  • Check for unexpected warnings in debug mode (nextflow run . -profile debug,test,docker --outdir <OUTDIR>).
  • Usage Documentation in docs/usage.md is updated.
  • Output Documentation in docs/output.md is updated.
  • CHANGELOG.md is updated.
  • README.md is updated (including new tool citations and authors/contributors).

Description

When --transcript_fasta and --additional_fasta are both provided without a pre-built pseudo-aligner index, the pipeline fails at quantification with a cryptic "missing transcripts" error. This happens because:

  • additional_fasta sequences are appended to the genome FASTA/GTF
  • But user-provided transcript_fasta is used as-is (without spike-ins) for building the index

Changes

  • Validation: Fail fast with clear error when building a pseudo-aligner index from transcript_fasta while additional_fasta is also set
  • Index-specific check: Only validates the relevant index for the selected pseudo-aligner (salmon_index for salmon, kallisto_index for kallisto)
  • Test fix: Kallisto test now sets additional_fasta = null since no kallisto index with spike-ins exists in test datasets
  • Documentation: Updated schema help text and usage docs with compatibility notes

Valid configurations

The combination is allowed when a pre-built index containing the spike-ins is provided:

# This will error (index built without spike-ins)
--transcript_fasta tx.fa --additional_fasta ercc.fa --pseudo_aligner salmon

# This works (pre-built index already has spike-ins)
--transcript_fasta tx.fa --additional_fasta ercc.fa --salmon_index /path/to/index

Closes #1450


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Copilot AI changed the title [WIP] Add validation error for incompatible transcript_fasta and additional_fasta params Add validation error for incompatible transcript_fasta and additional_fasta params Nov 25, 2025
Copilot AI requested a review from pinin4fjords November 25, 2025 21:26
@pinin4fjords pinin4fjords marked this pull request as ready for review November 25, 2025 21:27
@pinin4fjords pinin4fjords merged commit ac7a2e9 into fix/transcript-fasta-additional-fasta-validation Nov 25, 2025
@pinin4fjords pinin4fjords deleted the copilot/sub-pr-1632 branch November 25, 2025 21:27
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2 participants