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.
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.
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
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.