How to run a bulk RNA-seq differential expression analysis
Когда использовать: You have paired-end FASTQ, a reference, and a condition table.
Предварительные требования
- Skill installed — git clone https://github.com/jaechang-hits/SciAgent-Skills ~/.claude/skills/sciagent-skills
- Conda/mamba + compute — Use a Linux server or HPC; macOS fine for small datasets
Поток
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Plan the pipelinePlan a bulk RNA-seq DE workflow for 2 conditions x 3 reps with salmon + tximport + DESeq2. Output expected files per step.✓ Скопировано→ Step list with tools and intermediate outputs
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Run quantificationRun salmon quant for each sample; produce a script.✓ Скопировано→ Bash script with salmon invocations
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DE analysisLoad quants via tximport, run DESeq2, produce MA + volcano plots and a top-50 gene table.✓ Скопировано→ R script + output files
Итог: A DE table + plots you can hand to a PI or collaborator.
Подводные камни
- Mis-matched reference versus annotation GTF — Check release numbers explicitly; salmon and DESeq2 can silently run on mismatched IDs
- Low-count filtering removes biological signal — Use independent filtering or raise threshold gradually; don't blanket filter