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Low Input Whole-Genome Bisulfite Sequencing Using a Post-Bisulfite Adapter Tagging Approach

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Book cover DNA Methylation Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1708))

Abstract

The epigenetic mark 5-methylcytosine confers heritable regulation of gene expression that can be dynamically modulated during transitions in cell fate. With the development of high-throughput sequencing technologies, it is now possible to obtain comprehensive genome-wide maps of the mammalian DNA methylation landscape, but the application of these techniques to limited material remains challenging. Here, we present an optimized protocol to perform whole-genome bisulfite sequencing on low inputs (100–5000 somatic cells) using a post-bisulfite adapter tagging approach. In this strategy, bisulfite treatment is performed prior to library generation in order to both convert unmethylated cytosines and fragment DNA to an appropriate size. Then sequencing adapters are added by complementary strand synthesis using random tetramer priming, and libraries are subsequently amplified by PCR.

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Author information

Authors and Affiliations

  1. Epigenetics Programme, Babraham Institute, Babraham Research Campus, Cambridge, CB22 3AT, UK

    Julian R. Peat & Sébastien A. Smallwood

  2. Friedrich Miescher Institute for Biomedical Research, PO Box 3775, Maulbeerstrasse 66, CH-4058, Basel, Switzerland

    Sébastien A. Smallwood

Authors
  1. Julian R. Peat
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  2. Sébastien A. Smallwood
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Corresponding author

Correspondence to Sébastien A. Smallwood .

Editor information

Editors and Affiliations

  1. Laboratory for Epigenetics and Environment, Centre National de Recherche en Génomique Humaine, CEA—Institut de Biologie Francois Jacob, Evry, France

    Jörg Tost

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Peat, J.R., Smallwood, S.A. (2018). Low Input Whole-Genome Bisulfite Sequencing Using a Post-Bisulfite Adapter Tagging Approach. In: Tost, J. (eds) DNA Methylation Protocols. Methods in Molecular Biology, vol 1708. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7481-8_9

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  • DOI: https://doi.org/10.1007/978-1-4939-7481-8_9

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7479-5

  • Online ISBN: 978-1-4939-7481-8

  • eBook Packages: Springer Protocols

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