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Multiple Myeloma pp 179–191Cite as

A Rapid and Robust Protocol for Reduced Representation Bisulfite Sequencing in Multiple Myeloma

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1792))

Abstract

Reduced representation bisulfite sequencing (RRBS) is one of the most comprehensive yet economic ways of mapping whole genome DNA-methylation. Here, we have substantially modified the RRBS protocol by combining end-repair and A-tailing steps, and by introducing a bead-based method for rapid and easy size selection of the library molecules. The method has been optimized for myeloma clinical samples, where the input DNA concentration can be as low as 100 ng. The method developed can be accomplished in 3 days, including the initial overnight MspI enzyme digestion. Although the protocol has been optimized in myeloma samples, it is broadly applicable to any clinical sample, which is restricted by very low input DNA concentrations.

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Change history

  • 07 February 2019

    In Chapter 4, the author’s name Homer Adams III has been modified to Adams HC 3rd, as requested by him.

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Authors and Affiliations

  1. Department of Internal Medicine, Myeloma Institute, University of Arkansas for Medical Sciences, Little Rock, AR, USA

    Samrat Roy Choudhury & Brian A. Walker

Authors
  1. Samrat Roy Choudhury
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  2. Brian A. Walker
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Corresponding author

Correspondence to Brian A. Walker .

Editor information

Editors and Affiliations

  1. Janssen R&D, Spring House, PA, USA

    Christoph Heuck

  2. The Myeloma Institute, University of Arkansas for Medical Science, Little Rock, AR, USA

    Niels Weinhold

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Choudhury, S.R., Walker, B.A. (2018). A Rapid and Robust Protocol for Reduced Representation Bisulfite Sequencing in Multiple Myeloma. In: Heuck, C., Weinhold, N. (eds) Multiple Myeloma. Methods in Molecular Biology, vol 1792. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-7865-6_13

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  • DOI: https://doi.org/10.1007/978-1-4939-7865-6_13

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-7864-9

  • Online ISBN: 978-1-4939-7865-6

  • eBook Packages: Springer Protocols

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