Learner Profiles

This lesson is designed for researchers who want to perform a complete RNA-seq analysis using standard tools and reproducible workflows. Learners are typically graduate students, postdocs, research staff, or faculty working with sequencing-based gene expression data.

Background and Experience

  • Familiarity with basic biological concepts including genes, transcripts and expression
  • Some exposure to the command line (navigating directories, running commands)
  • Little or no prior experience with RNA-seq data analysis
  • No requirement for programming experience beyond running provided scripts

Motivations

  • Analyze RNA-seq data from their own experiments
  • Learn standard QC, alignment, quantification and differential expression workflows
  • Understand how to interpret results and generate publication-ready figures
  • Gain exposure to reproducible analysis approaches on HPC systems

Needs and Goals

  • Know how to evaluate read quality
  • Learn how to align reads and generate count matrices
  • Perform differential expression analysis using established statistical frameworks
  • Understand normalization, modeling assumptions and interpretation of outputs
  • Learn best practices for organizing projects and making workflows reproducible

Challenges

  • Managing multiple tools and file formats
  • Understanding statistical concepts such as dispersion, FDR and normalization
  • Navigating HPC environments and resource allocation
  • Interpreting QC metrics and identifying technical artifacts

Computing Requirements

  • Access to a Unix-like shell environment (local or cluster)
  • Ability to run command-line tools and R-based analysis
  • Internet browser for visual summaries and documentation access