RNA-seq Analysis 2024

Welcome

Welcome to RNA-seq Analysis 2024.

The course schedule can be found here

Pre-Workshop Preparation

Pre-work including programs to install can be found here

AWS Module

Connecting and properly using a cloud computing cluster at the CBW here


Class Photo

RNA-seq Analysis Class of 2024


Course Materials

Day 1

Welcome

Nia Hughes

Module 1

Module 1 Lecture Recording

Module 1 Lecture Slides - Intro
Module 1 Lecture Slides - FASTA/FASTQ/GTF
Module 1 Lecture Slides - Indexing

Module 1 Lab

Introduction to Inputs
Reference Genomes
Annotations
Indexing
Data
Data QC

Module 2

Module 2.1 Lecture Recording
Module 2.2 Lecture Recording

Module 2 Lecture Slides - Alignment
Module 2 Lecture Slides - Alignment vs Assembly vs Kmer
Module 2 Lecture Slides - SAM/BAM/BED
Module 2 Lecture Slides - IGV
Module 2 Lecture Slides - Alignment QC

Module 2 Lab

Introduction to Alignment
Adapter Trim
Alignment
IGV
Alignment Visualization Preparation
Alignment Visualization IGV
Alignment Visualization Allele Expression
Alignment QC

Optional Team Assignment

Team Assignment - Alignment

Complete Part I - Obtaining the dataset & reference files and Part II - Data Preprocessing.

Day 2

Module 3

Module 3 Lecture Recording

Module 3 Lecture Slides

Module 3 Lab

Introduction to Expression Analysis
Expression Analysis with Stringtie and htseq-count

Optional Team Assignment

Team Assignment - Alignment (Continued)

Complete Part III - Alignment and Part IV - Post-alignment QC & IGV Visualization.

Team Assignment - Expression and DE

Complete Part I - Expression Estimation.

Day 3

Module 3 Lab (Continued)

Differential Expression with Ballgown
Differential Expression with DESeq2
DE Visualization with Ballgown
DE Visualization with DESeq2
DE Visualization Advanced
DE Pathway Analysis
Batch Correction - Preparation
Batch Correction Analysis

Post-Workshop Materials

AMI

We have made our AWS AMI (Amazon Machine Image) publicly available. To launch your own instance, follow the instructions provided by Amazon on how to launch an EC2 instance from a custom Amazon Machine Image. Please note that you will need an AWS account to proceed, and that you will need to upload the CourseData files yourself.

Here are the details of the AMI:

  • AWS Region: us-east-1 (N. Virgina)
  • AMI type: public image
  • AMI name: CBW_RNA_240613
  • AMI ID: ami-0216fae97002bb976

If you want to create and activate a new AWS account, please follow the instructions provided by Amazon.