Generation of Whole Genome Bisulfite Sequencing Libraries for Comprehensive DNA Methylome Analysis

Dulce B Vargas-Landin, Jahnvi Pflüger, Ryan Lister

Research output: Chapter in Book/Conference paperChapterpeer-review

3 Citations (Scopus)

Abstract

Whole genome bisulfite sequencing (WGBS) enables the detection of DNA methylation at single base-pair resolution. The treatment of DNA with sodium bisulfite allows the discrimination of methylated and unmethylated cytosines, but the power of this technology can be limited by the input amounts of DNA and the length of DNA fragments due to DNA damage caused by the desulfonation process. Here, we describe a WGBS library preparation protocol that minimizes the loss and damage of DNA, generating high quality libraries amplified with fewer PCR cycles, and hence data with fewer PCR duplicates, from lower amounts of input material. Briefly, genomic DNA is sheared, end-repaired, 3'-adenylated, and ligated to adaptors with fewer cleanup steps in between, minimizing DNA loss. The adapter-ligated DNA is then treated with sodium bisulfite and amplified with few PCR cycles to reach the yield needed for sequencing.

Original languageEnglish
Title of host publicationEpigenome Editing
Subtitle of host publicationMethods and Protocols
Place of PublicationNew York
PublisherSpringer
Pages291-298
Number of pages8
ISBN (Electronic)9781493977741
ISBN (Print)9781493977734
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1767
ISSN (Print)1064-3745

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