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Tagmentation-based whole-genome bisulfite sequencing

by Radlwimmer, Bernhard , Gu, Lei , Wolf, Stephan , Shendure, Jay , Wang, Wei , Plass, Christoph , Weichenhan, Dieter , Wang, Qi , Eils, Roland , Hovestadt, Volker , Bähr, Marion , Adey, Andrew

in 631/1647/2210/2213 / 631/1647/514/2254 / 631/208/176/1988 / 631/208/212/177 / Adapters / Analysis / Analytical Chemistry / Animals / Bar codes / Biological Techniques / Bisulfite / Computational Biology/Bioinformatics / CpG islands / Cytosine / Deoxyribonucleic acid / DNA / DNA Methylation / DNA sequencing / Epigenesis, Genetic / Epigenetics / Genomes / Genomics - methods / Humans / Life Sciences / Methylation / Mice / Microarrays / Nucleotide sequence / Organic Chemistry / Protocol / Sequence Analysis, DNA - methods / Sulfites / Sulfites - chemistry / Transposase

2013

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Tagmentation-based whole-genome bisulfite sequencing

by Radlwimmer, Bernhard , Gu, Lei , Wolf, Stephan , Shendure, Jay , Wang, Wei , Plass, Christoph , Weichenhan, Dieter , Wang, Qi , Eils, Roland , Hovestadt, Volker , Bähr, Marion , Adey, Andrew

2013

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Tagmentation-based whole-genome bisulfite sequencing

by Radlwimmer, Bernhard , Gu, Lei , Wolf, Stephan , Shendure, Jay , Wang, Wei , Plass, Christoph , Weichenhan, Dieter , Wang, Qi , Eils, Roland , Hovestadt, Volker , Bähr, Marion , Adey, Andrew

2013

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Journal Article

Tagmentation-based whole-genome bisulfite sequencing

Radlwimmer, Bernhard,

Gu, Lei,

Wolf, Stephan,

Shendure, Jay,

Wang, Wei,

Plass, Christoph,

Weichenhan, Dieter,

Wang, Qi,

Eils, Roland,

Hovestadt, Volker,

Bähr, Marion,

Adey, Andrew

2013

Overview

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 Tn 5 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 2 d.

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Publisher

Nature Publishing Group UK,Nature Publishing Group

Subject

/ Analytical Chemistry