gitwtfhub

wtf is 001_rnaseq_downstream_pipeline?

lamarcklab/001_rnaseq_downstream_pipeline — explained in plain English

Analysis updated 2026-05-18

53RAudience · researcherComplexity · 3/5Setup · moderate

TL;DR

A step by step R pipeline that turns RNA sequencing gene counts into findings about which genes and pathways changed between two conditions.

Mindmap

mindmap
  root((RNAseq Pipeline))
    What it does
      Analyzes gene counts
      Finds changed genes
      Pathway enrichment
    Tech stack
      R
      DESeq2
      Bioconductor
    Stages
      QC
      Differential expression
      GO KEGG enrichment
      GSEA
    Use cases
      Find affected pathways
      Learn RNAseq workflow
      Reproduce plots
    Audience
      Biology researchers
      Bioinformatics students

Code map

Detail Auto

An interactive map of this repo's files and how they connect — its source is parsed live in your browser. Click Visualize to build it.

filefunction / class

Why would anyone build with this?

REASON 1

Analyze RNA sequencing count data to find genes that turned on or off between two conditions.

REASON 2

Run GO and KEGG pathway enrichment to see which biological processes a treatment affects.

REASON 3

Learn a working example of a full RNA-seq downstream analysis workflow in R.

REASON 4

Reproduce published-style volcano plots, bubble charts, and enrichment networks from gene expression data.

What's in the stack?

RDESeq2Bioconductor

How it stacks up

lamarcklab/001_rnaseq_downstream_pipelinebrendangregg/perfmodelshadley/mylittlepony
Stars537218
LanguageRRR
Last pushed2014-12-052018-11-09
MaintenanceDormantDormant
Setup difficultymoderatemoderateeasy
Complexity3/53/51/5
Audienceresearcherops devopsdeveloper

Figures from each repo's GitHub metadata at analysis time.

How do you spin it up?

Difficulty · moderate Time to first run · 1h+

Requires installing several specific R and Bioconductor packages and editing hardcoded file paths.

Wtf does this do

This repository is a step by step recipe, written in the R programming language, for analyzing RNA sequencing data after the raw counting has already been done. RNA sequencing is a lab method that measures which genes are active in a cell or tissue sample. This project takes that starting data and walks through the process of figuring out which genes changed, and what those changes might mean biologically. The pipeline runs through seven stages in order: quality control checks, finding differentially expressed genes (genes that turned on or off between two conditions), converting gene ID formats so different databases can talk to each other, and then three kinds of enrichment analysis that group genes into known biological pathways to look for patterns. Each stage lives in its own folder with its own scripts and its own README explaining that step. To make the example realistic without using anyone's private data, the project uses a public, published dataset from a 2014 scientific paper. That dataset compares airway muscle cells that were treated with a steroid drug against cells that were not treated, and the pipeline demonstrates how to find which genes and pathways the drug affects. Running it requires installing R and a handful of specific R packages, then running a setup script to prepare the starting files, then running each stage's scripts in order since later steps depend on the output of earlier ones. The scripts currently point to a specific folder path on the original author's computer, so a new user needs to edit that path before running anything. This is best suited for people who already work with RNA sequencing data and know some R, such as biology researchers or bioinformatics students, rather than complete beginners to coding or to genomics.

Yoink these prompts

Prompt 1
Walk me through what each of the 7 stages in this RNA-seq pipeline does and why the order matters.
Prompt 2
Help me edit the absolute file paths in these R scripts so the pipeline runs on my own computer.
Prompt 3
Explain what GO enrichment and KEGG enrichment are and how to interpret the plots this pipeline produces.
Prompt 4
Show me how to swap the airway demo dataset for my own RNA-seq count matrix in this pipeline.
Prompt 5
What R packages do I need to install before I can run this RNA-seq pipeline end to end?

Frequently asked questions

wtf is 001_rnaseq_downstream_pipeline?

A step by step R pipeline that turns RNA sequencing gene counts into findings about which genes and pathways changed between two conditions.

What language is 001_rnaseq_downstream_pipeline written in?

Mainly R. The stack also includes R, DESeq2, Bioconductor.

How hard is 001_rnaseq_downstream_pipeline to set up?

Setup difficulty is rated moderate, with roughly 1h+ to a first successful run.

Who is 001_rnaseq_downstream_pipeline for?

Mainly researcher.

View the repo → Decode another repo

This repo across BitVibe Labs

Don't trust strangers blindly. Verify against the repo.