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A beginner’s guide to bulk RNA-Seq analysis

Laura Saba, PhD

Materials from Webinar

Webinar slides

General Papers

Survey of best practices - Conesa et al 2016

Introduction to RNA-Seq - Kukurba and Montgomery 2015

Key References from Presentation

Technical Overview

Overview of RNA-Seq - Pepke et al 2009

RNA Separation - Mattick & Makunin 2006

RNA Separation - Genomes 2002

RNA Separation - Kukurba and Montgomery 2015

Library Preparation - Corney and Basturea 2013

Link to Illumina Video - Illumina Sequencing by Synthesis

Experimental Design

Sequencing Depth - Sims et al 2014

ERCC Synthetic Spike-Ins - ThermoFisher

Transcriptome Profiling

Popular Pipelines

Trimming

Read Alignment

Transcript Discovery

Quantification

Differential Expression

DESeq2

False Discovery Rate

Alternative splicing - MISO

Alternative polyadenylation - DaPars

Additional References

Influence of RNA extraction - Sultan et al 2014

RNA-Seq Power Analyses/Sample Size Calculations - Ching et al 2014

RNA-Seq Power Analyses/Sample Size Calculations - Bi and Lui 2016

RNA-Seq Power Analyses/Sample Size Calculations - Busby et al 2013

ERCC Synthetic Spike-Ins for RNA-Seq - Jiang et al 2011