Bioinformatics RNA-seq Analysis: From Raw Reads to Biological Insights

Master complete RNA-seq analysis workflows from quality control to biological interpretation. Learn industry-standard tools, differential expression analysis, and functional enrichment using real datasets and cloud platforms.

Workshop Description

This intensive 4-day workshop provides researchers with comprehensive training in RNA-seq data analysis from raw sequencing reads to biological interpretation. Designed for biologists and researchers with basic computational skills, this course covers the complete RNA-seq pipeline using industry-standard tools and best practices.

You'll master essential bioinformatics concepts including quality control, read alignment, quantification, differential expression analysis, and functional enrichment. The workshop emphasizes hands-on experience with real datasets, using command-line tools, R/Bioconductor packages, and cloud-based platforms to ensure reproducible and publication-ready analyses.

By the end of this workshop, you'll be confident performing end-to-end RNA-seq analysis, interpreting results in biological context, creating publication-quality figures, and applying these skills to your own research datasets. All software, datasets, and analysis scripts are provided, with lifetime access to materials and continued support.

Instructor

Dr Victor Gambarini

Course Fee

$449.0

Maximum Seats

20

Duration

4 half-days

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Time
05:00 - 09:00
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A comprehensive 4-day journey through modern RNA-seq analysis workflows and interpretation

Format: Each day is approximately 4 hours with hands-on analysis using real RNA-seq datasets on cloud platforms

Prerequisites: Basic biology background and familiarity with R or Python recommended. Some command-line experience helpful but not required. We'll provide quick tutorials for essential skills

What you'll learn: Complete RNA-seq pipeline from raw reads to biological insights, quality control, alignment strategies, quantification methods, statistical analysis, and functional interpretation

Materials provided: All datasets, analysis scripts, reference materials, and cloud computing access. Lifetime access to updated protocols and continued community support

Day 1: RNA-seq Fundamentals & Quality Control

Understand RNA-seq technology and perform essential quality assessments

RNA-seq Overview (45 min): Sequencing technologies, experimental design considerations, single-end vs paired-end reads, library preparation methods, and common experimental pitfalls

File Formats & Data Structure (30 min): FASTQ, SAM/BAM, GTF/GFF formats, understanding sequencing metadata, and organizing bioinformatics projects

Quality Control with FastQC (1 hr): Assessing read quality, identifying common issues (adapter contamination, low quality regions), interpreting quality metrics, and batch processing multiple samples

Read Preprocessing (1 hr): Trimming adapters and low-quality bases with Trimmomatic, filtering strategies, and quality improvement verification

Introduction to Command Line (45 min): Essential Unix commands for bioinformatics, file manipulation, and basic scripting for reproducible workflows

Day 2: Read Alignment & Quantification

Map reads to reference genomes and quantify gene expression

Reference Genomes & Annotation (30 min): Choosing appropriate references, understanding genome builds, annotation files (GTF/GFF), and handling model vs non-model organisms

Alignment Strategies (45 min): Splice-aware aligners (STAR, HISAT2), alignment parameters, handling splice junctions, and alignment quality metrics

Hands-on Alignment (1.5 hrs): Indexing reference genomes, running STAR alignment, understanding output files, and assessing alignment quality with samtools and RSeQC

Gene Quantification (1 hr): Count-based methods (htseq-count, featureCounts), abundance estimation (Salmon, Kallisto), and choosing quantification strategies

Quality Assessment (1 hr): MultiQC for comprehensive quality reports, identifying problematic samples, and documenting analysis decisions

Day 3: Differential Expression Analysis

Statistical analysis and identification of differentially expressed genes

Introduction to R/Bioconductor (30 min): Setting up R environment, essential Bioconductor packages (DESeq2, edgeR, limma), and data import strategies

Experimental Design & Statistics (45 min): Understanding biological vs technical replicates, power analysis, batch effects, and statistical models for RNA-seq

DESeq2 Analysis Pipeline (2 hrs): Data preprocessing, normalization methods, dispersion estimation, statistical testing, multiple testing correction, and results interpretation

Visualization & Exploration (1 hr): PCA plots, heatmaps, volcano plots, MA plots using ggplot2 and specialized packages, and identifying patterns in expression data

Advanced Considerations (15 min): Handling complex experimental designs, time-course experiments, and multi-factor analyses

Day 4: Functional Analysis & Biological Interpretation

Transform gene lists into biological insights and publication-ready results

Gene Set Enrichment (1.5 hrs): Gene Ontology (GO) analysis, KEGG pathway enrichment, using clusterProfiler and enrichR, statistical considerations, and avoiding common pitfalls

Advanced Functional Analysis (1 hr): Gene Set Enrichment Analysis (GSEA), pathway visualization with pathview, network analysis, and integrating multiple data types

Visualization & Reporting (1 hr): Creating publication-quality figures, R Markdown reports for reproducible analysis, and integrating results across analysis steps

Case Study & Integration (30 min): Complete analysis walkthrough with real biological dataset, from hypothesis to biological conclusions, troubleshooting common issues

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