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The aim of this course is to prepare and analyze "bisulfite-free" methylome libraries for Illumina sequencing. We will use a novel enzymatic approach, to detect 5-methyl cytosines (5mC's), that results in longer reads and higher coverage of genomic features at lower minimum coverage thresholds when compared to Illumina bisulfite libraries.
The course participants will be guided through the novel workflow for bisulfite-free library generation. Illumina libraries will be produced using the 50 ng control genomic DNA. Attendees will benefit from hands on experience and gain insight into the method as they progress through the protocol. In addition to the control DNA, participants can also provide a maximum of 2 samples for library preparation, with a required minimum of 100 ng DNA per sample. Libraries will be sequenced, and bioinformatics analysis of a dataset will be discussed.
This course is directed towards wet-lab biologists, scientists and technicians who want to apply the bisulfite-free method for detecting 5mC's to their samples. Participants will learn the best practice of library preparation and data analysis. Knowledge of next generation sequencing technology is desirable but not essential.
- Enzymatic detection of 5mC's
- Introduction to methylome data analysis
Preparation of methylome libraries using an enzymatic approach. Introduction to data analysis of these libraries.