ABI Bioinformatics Guide 2024
  • INTRODUCTION
    • How to use the guide
  • MOLECULAR BIOLOGY
    • The Cell
      • Cells and Their Organelles
      • Cell Specialisation
      • Quiz 1
    • Biological Molecules
      • Carbohydrates
      • Lipids
      • Nucleic Acids (DNA and RNA)
      • Quiz 2
      • Proteins
      • Catalysis of Biological Reactions
      • Quiz 3
    • Information Flow in the Cell
      • DNA Replication
      • Gene Expression: Transcription
      • Gene Expression: RNA Processing
      • Quiz 4
      • Chromatin and Chromosomes
      • Regulation of Gene Expression
      • Quiz 5
      • The Genetic Code
      • Gene Expression: Translation
    • Cell Cycle and Cell Division
      • Quiz 6
    • Mutations and Variations
      • Point mutations
      • Genotype-Phenotype Interactions
      • Quiz 7
  • PROGRAMMING
    • Python for Genomics
    • R programming (optional)
  • STATISTICS: THEORY
    • Introduction to Probability
      • Conditional Probability
      • Independent Events
    • Random Variables
      • Independent, Dependent and Controlled Variables
    • Data distribution PMF, PDF, CDF
    • Mean, Variance of a Random Variable
    • Some Common Distributions
    • Exploratory Statistics: Mean, Median, Quantiles, Variance/SD
    • Data Visualization
    • Confidence Intervals
    • Comparison tests, p-value, z-score
    • Multiple test correction: Bonferroni, FDR
    • Regression & Correlation
    • Dimentionality Reduction
      • PCA (Principal Component Analysis)
      • t-SNE (t-Distributed Stochastic Neighbor Embedding)
      • UMAP (Uniform Manifold Approximation and Projection)
    • QUIZ
  • STATISTICS & PROGRAMMING
  • BIOINFORMATICS ALGORITHMS
    • Introduction
    • DNA strings and sequencing file formats
    • Read alignment: exact matching
    • Indexing before alignment
    • Read alignment: approximate matching
    • Global and local alignment
  • NGS DATA ANALYSIS & FUNCTIONAL GENOMICS
    • Experimental Techniques
      • Polymerase Chain Reaction
      • Sanger (first generation) Sequencing Technologies
      • Next (second) Generation Sequencing technologies
      • The third generation of sequencing technologies
    • The Linux Command-line
      • Connecting to the Server
      • The Linux Command-Line For Beginners
      • The Bash Terminal
    • File formats, alignment, and genomic features
      • FASTA & FASTQ file formats
      • Basic Unix Commands for Genomics
      • Sequences and Genomic Features Part 1
      • Sequences and Genomic Features Part 2: SAMtools
      • Sequences and Genomic Features Part 3: BEDtools
    • Genetic variations & variant calling
      • Genomic Variations
      • Alignment and variant detection: Practical
      • Integrative Genomics Viewer
      • Variant Calling with GATK
    • RNA Sequencing & Gene expression
      • Gene expression and how we measure it
      • Gene expression quantification and normalization
      • Explorative analysis of gene expression
      • Differential expression analysis with DESeq2
      • Functional enrichment analysis
    • Single-cell Sequencing and Data Analysis
      • scRNA-seq Data Analysis Workflow
      • scRNA-seq Data Visualization Methods
  • FINAL REMARKS
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  • DESeq2 tutorial #1 by Griffith lab
  • DESeq2 tutorial #2 by Altuna Akalin

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  1. NGS DATA ANALYSIS & FUNCTIONAL GENOMICS
  2. RNA Sequencing & Gene expression

Differential expression analysis with DESeq2

PreviousExplorative analysis of gene expressionNextFunctional enrichment analysis

Last updated 5 months ago

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Differential gene expression analysis is a technique used to identify genes that are differentially expressed, or turned on or off, between two or more biological conditions or samples. This can be used to identify genes that are specifically involved in a particular process or disease, or to understand the underlying molecular mechanisms of a biological system.

Once the differentially expressed genes have been identified, researchers can use various techniques, such as gene ontology analysis and pathway analysis, to further understand the biological functions and pathways in which these genes are involved. This information can be used to gain insights into the underlying mechanisms of a biological process or disease, and may ultimately lead to the development of new treatments or therapies.

There are several tools used to identify genes differentially expressed between different conditions or groups of samples. Here, we will explore DESeq2 (Differential Expression analysis for Sequencing). It uses statistical methods to analyze RNA-seq data and identify genes that are differentially expressed between two or more conditions or samples. It takes into account various sources of variability in the data, such as batch effects and technical noise, to accurately identify differentially expressed genes.

We are offering you two tutorials to run DESeq2 on two different samples.

DESeq2 tutorial #1 by Griffith lab

Use your local R Studio and replicate the pipeline described in this tutorial.

DESeq2 tutorial #2 by Altuna Akalin

You can also try out this tutorial, where you will find additional explanation for the pipeline. Note that in order to be able to access the dataset for this tutorial you'll need to have run the previous R practicals of this textbook subchapter.

Differential expression with DEseq2Griffith Lab
8.3 Gene expression analysis using high-throughput sequencing technologies | Computational Genomics with R
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