How to run a bulk RNA-seq differential expression analysis
使うタイミング: You have paired-end FASTQ, a reference, and a condition table.
前提条件
- 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
フロー
-
Plan the pipelinePlan a bulk RNA-seq DE workflow for 2 conditions x 3 reps with salmon + tximport + DESeq2. Output expected files per step.✓ コピーしました→ Step list with tools and intermediate outputs
-
Run quantificationRun salmon quant for each sample; produce a script.✓ コピーしました→ Bash script with salmon invocations
-
DE analysisLoad quants via tximport, run DESeq2, produce MA + volcano plots and a top-50 gene table.✓ コピーしました→ R script + output files
結果: A DE table + plots you can hand to a PI or collaborator.
注意点
- 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