Course Overview

The goal of this course is to teach the fundamentals of NGS library preparation for single-cell transcriptome analysis using the Illumina & Bio-Rad Single-Cell Sequencing Solution. We will start by a general introduction on the topic, including the Illumina NGS technology, Biorad droplet partitioning technology and the library preparation workflow. The hands-on part begins with cell generation, followed by the preparation of transcriptome libraries from single-cells, with a focus on the critical steps and potential pitfalls and best practices of the protocol. Finally, we will quantify and quality control the prepared libraries, before sequencing the libraries on a NextSeq instrument. The last day will be dedicated to discussing and running-through solutions for single-cell data analysis.

Audience

This course is directed towards biologists, scientists and technicians who want to apply single-cell transcriptome sequencing and learn the basics of library preparation. Knowledge of next generation sequencing technology and cell culture is desirable but not essential.

Modules/Resources

During this course we will focus on the following aspects:

• Single-Cell preparation
• Best practices and tips and tricks for preparation 3’-tagged RNA-Seq libraries from single cells for whole transcriptome gene profiling analysis
• Best practices of sequencing library validation
• Introduction to single cell transcriptome data analysis

Learning Outcomes

After this course you should be able to:

• Prepare libraries from cell cultures for single-cell whole transcriptome analysis
• Apply quality controls and library quantification
• Know the basics of single-cell transcriptome data analysis

This course is co-organised with Illumina & BioRad.