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

53 Citations (Scopus)

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 2013
Externally publishedYes

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Genes
Genome
Methylation
DNA
Cytosine
DNA Methylation
Bioinformatics
Computational Biology
DNA Sequence Analysis
Epigenomics
Libraries
Disease Progression
Carbon
hydrogen sulfite
Costs and Cost Analysis
Costs

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Wang, Q., Gu, L., Adey, A., Radlwimmer, B., Wang, W., Hovestadt, V., ... Weichenhan, D. (2013). Tagmentation-based whole-genome bisulfite sequencing. Nature Protocols, 8(10), 2022-2032. https://doi.org/10.1038/nprot.2013.118

Tagmentation-based whole-genome bisulfite sequencing. / Wang, Qi; Gu, Lei; Adey, Andrew; Radlwimmer, Bernhard; Wang, Wei; Hovestadt, Volker; Bähr, Marion; Wolf, Stephan; Shendure, Jay; Eils, Roland; Plass, Christoph; Weichenhan, Dieter.

In: Nature Protocols, Vol. 8, No. 10, 10.2013, p. 2022-2032.

Research output: Contribution to journalArticle

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, vol. 8, no. 10, pp. 2022-2032. https://doi.org/10.1038/nprot.2013.118
Wang Q, Gu L, Adey A, Radlwimmer B, Wang W, Hovestadt V et al. Tagmentation-based whole-genome bisulfite sequencing. Nature Protocols. 2013 Oct;8(10):2022-2032. https://doi.org/10.1038/nprot.2013.118
Wang, Qi ; Gu, Lei ; Adey, Andrew ; Radlwimmer, Bernhard ; Wang, Wei ; Hovestadt, Volker ; Bähr, Marion ; Wolf, Stephan ; Shendure, Jay ; Eils, Roland ; Plass, Christoph ; Weichenhan, Dieter. / Tagmentation-based whole-genome bisulfite sequencing. In: Nature Protocols. 2013 ; Vol. 8, No. 10. pp. 2022-2032.
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