The goal of this hands-on course is to teach the fundamentals of RNA and DNA library preparation for next generation sequencing applications. RNA depletion and enrichment strategies for different populations of RNA will be explained and users will learn about the advantages of RNA-Seq compared to other methods such as Sanger sequencing and microarrays.
In addition, library preparation for (low input) DNA samples will be performed. Users will also learn about sample QC and library quantification that enable the preparation of high-quality, low-biased DNA libraries.
Participants are highly encouraged to bring their own RNA and DNA samples to this hands-on course.
The bioinformatics part of the course will cover tools for RNA-seq data analysis using open-source software. Computers for hands-on exercises will be provided along with demo data sets.
No prior NGS or coding experience is required to participate in this course. This course was developed for biologists who are curious to learn more about NGS and data analysis as the intended audience.
- Major strategies to prepare NGS libraries.
- RNA library preparation with improved sequencing coverage and uniformity of GC-rich and low abundant transcripts (rRNA depletion, mRNA capture, no RNA enrichment).
- Construction of high-quality DNA libraries from low-input amounts and challenging samples.
- Library quantification with KAPA Library Quantification (qPCR) to reliable pool NGS libraries for capture or flow cell amplification.
- Tools for RNA-seq data analysis and biological interpretation
- DNA and RNA library preparation from a variety of sample types using KAPA Hyper/HyperPlus technology.
- How to properly quantify and set up QC for NGS libraries to accurately, reliably, and reproducible guarantee flow cell amplification for Illumina® sequencers.
- Best practices, tips and tricks for NGS library preparation, including challenging sample types (e.g. FFPET).
- Assess the quality of RNA-seq data and complete basic statistical tests on NGS data
This course is co-organised with Roche.