RNAseq Laboratory Workshop


This training course includes presentations on technology theory and practical experience with the entire laboratory sample prep workflow. Experienced trainers will lead you through using TapeStationand Miseqequipment. By the end of this training, participants will be able to complete the Illumina library preparation process and set up a sequencing run using Illumina MiseqControl Software.

Table of Contents

  1. Pre-course Materials
  2. Presentation Slides

Pre-course Materials

You should have received an email including the agenda and pre-course materials for this workshop.

Illumina Sequencing Overview

Introduction to MiSeq and the NextSeq

Best Practices for TruSeq Stranded mRNA

Overview of Illumina NGS Library Preparation Workflow Overview of Illumina NGS library preparation workflows include understanding the basics of NGS chemistry, library multiplexing and the difference between the library preparations in all NSG methods such as whole genome sequencing (WGS), targeted sequencing (16s sequencing or exome seq), RNA sequencing, Methylation sequencing and ChIP sequencing, etc.

Understanding Illumina SBS TechnologySequencing by Synthesis (SBS) technology uses four fluorescently labeled nucleotides to sequence the tens of millions of clusters on the flow cell surface in parallel. During each sequencing cycle, a single labeled dNTP is added to the nucleic acid chain, after dNTP incorporation, the fluorescent dye is identified through laser excitation and imaging, then enzymatically cleaved to allow the next round of incorporation. Base calls are made directly from signal intensity measurements during each cycle.

Presentation Slides

Download the slides here:

Introduction to the USF Genomics Core and iLab System by Min Zhang

Illumina Sequencing Overview by Ryan Gentry (OLD)

Understanding Illumina sequencing technology (SBS) by Meredith Millis

Library Preparation for Next-Gen Sequencing by Min Zhang

Library QC, Trouble-shooting, Sequencing Data QC and Illumina BaseSpace Tour by Min Zhang

Illumina BaseSpace Tour and Sequencing data QC by Matt Angel

Sequencing set up and Illumina Customer Resources Overview by Matt Mercurio