Tagmentation-based whole-genome bisulfite sequencing

被引：127

作者：

Wang, Qi ^{[1
]}

Gu, Lei ^{[1
,2
]}

Adey, Andrew ^{[3
]}

Radlwimmer, Bernhard ^{[4
]}

Wang, Wei ^{[4
]}

Hovestadt, Volker ^{[4
]}

Baehr, Marion ^{[2
]}

Wolf, Stephan ^{[5
]}

Shendure, Jay ^{[3
]}

Eils, Roland ^{[1
]}

Plass, Christoph ^{[2
]}

Weichenhan, Dieter ^{[2
]}

机构：

[1] German Canc Res Ctr, Heidelberg, Germany

[2] DKFZ, Div Epigen & Canc Risk Factors, Heidelberg, Germany

[3] Univ Washington, Dept Genome Sci, Seattle, WA USA

[4] DKFZ, Div Mol Genet, Heidelberg, Germany

[5] DKFZ, Core Facil, Heidelberg, Germany

来源：

NATURE PROTOCOLS | 2013年 / 8卷 / 10期

基金：

美国国家科学基金会;

关键词：

PRIMORDIAL GERM-CELLS; DNA METHYLATION; CYTOSINE METHYLATION; LOW-INPUT; WIDE; LIBRARIES;

D O I：

10.1038/nprot.2013.118

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Epigenetic modifications such as carbon 5 methylation of the cytosine base in a CpG dinucleotide context are involved in the onset and progression of human diseases. A comprehensive understanding of the role of genome-wide DNA methylation patterns, the methylome, requires quantitative determination of the methylation states of all CpG sites in a genome. So far, analyses of the complete methylome by whole-genome bisulfite sequencing (WGBS) are rare because of the required large DNA quantities, substantial bioinformatic resources and high sequencing costs. Here we describe a detailed protocol for tagmentation-based WGBS (T-WGBS) and demonstrate its reliability in comparison with conventional WGBS. In T-WGBS, a hyperactive Tn5 transposase fragments the DNA and appends sequencing adapters in a single step. T-WGBS requires not more than 20 ng of input DNA; hence, the protocol allows the comprehensive methylome analysis of limited amounts of DNA isolated from precious biological specimens. The T-WGBS library preparation takes 2d.

引用

页码：2022 / 2032

页数：11

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