NGS – Library construction and quality control

2 days NGS – Library construction and quality control

Target audience: Beginners to Medium experienced NGS users

Entrance qualifications: Basic Molecular biology or similar

Description: This course gives an introduction to massively parallel sequencing (also called Next Generation Sequencing, NGS), and its many applications. The course consists of a theoretical part, which will focus on considerations for the NGS experiment design, the different sequencing platforms, quality control of samples, library preparation techniques, and quantification of libraries for sequencing. The course also includes practical parts where the participants will prepare libraries and perform quality control and compare libraries.

The course contains:

Introduction to NGS:

  • The history of DNA sequencing
  • What are the advantages with NGS? Disadvantages?
  • What is the difference between microarray and NGS?

Applications:

  • What are the possibilities for analysis of the genome? Transcriptome? Epigenome? Metagenome?
  • RNA sequencing; investigating differential expression, splice variants, novel transcripts and much more.
  • DNA sequencing; finding Single-Nucleotide Polymorphisms (SNPs), Copy Number Variants (CNVs), structural variations and more.
  • NGS for epigenetic studies; analyzing the methylation state of DNA or discovering protein-binding sites in DNA.

Sample preparation and quality control:

  • How to prepare samples for NGS.
  • Which starting material can be used for NGS?
  • Which quality controls are included in the experimental workflow?
  • How to choose the right library prep protocol.
  • Practical experiment to learn the basics of library preparation and quality control.

Instrumentation:

  • Which platforms are available?
  • How do they work?
  • Which platform is suitable for my experiment?

Experimental design:

  • What to think about when designing your NGS experiment.
  • How much to sequence?
  • How to calculate coverage and depth?
  • How many samples can I multiplex?