bio-workflows-rnaseq-to-de
Version Compatibility
Reference examples tested with: DESeq2 1.42+, tximport 1.30+, apeglm 1.24+, STAR 2.7.11+, Salmon 1.10+, Subread/featureCounts 2.0.2+ (--countReadPairs added in 2.0.2), fastp 0.23+, ggplot2 3.5+ (kallisto 0.50+ as a Salmon alternative)
Before using code patterns, verify installed versions match. If versions differ:
- R:
packageVersion('<pkg>')then?function_nameto verify parameters - CLI:
<tool> --versionthen<tool> --helpto confirm flags
If code throws ImportError, AttributeError, or TypeError, introspect the installed package and adapt the example to match the actual API rather than retrying.
Note: Salmon selective alignment is default since 1.0 (the historical --validateMappings is now a no-op); DESeqDataSetFromTximport carries the average-transcript-length offset automatically; lfcShrink(type='apeglm') requires coef to name a resultsNames(dds) coefficient and DROPS the stat column. Confirm these in-tool before quoting.
RNA-seq to Differential Expression Workflow
"Find differentially expressed genes from my RNA-seq FASTQ files" -> Chain QC/trim, decoy-aware quantification, tximport gene-level collapse, a count-based DE test, shrinkage, and visualization into one annotated DE table.
- CLI + R: fastp -> (salmon | STAR + featureCounts) -> tximport -> DESeq2/edgeR/limma-voom -> lfcShrink -> VST/volcano
This is a workflow skill: it owns the chaining decisions and hand-offs, not the internals of any one step. Every step below cross-references the component skill that teaches its mechanism.