Tagmentation-based whole-genome bisulfite sequencing

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

Research output: Contribution to journalArticle

63 Scopus citations

Abstract

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 2 d.

Original languageEnglish (US)
Pages (from-to)2022-2032
Number of pages11
JournalNature protocols
Volume8
Issue number10
DOIs
StatePublished - Oct 1 2013

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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    Wang, Q., Gu, L., Adey, A., Radlwimmer, B., Wang, W., Hovestadt, V., Bähr, M., Wolf, S., Shendure, J., Eils, R., Plass, C., & Weichenhan, D. (2013). Tagmentation-based whole-genome bisulfite sequencing. Nature protocols, 8(10), 2022-2032. https://doi.org/10.1038/nprot.2013.118