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Tagmentation-Based Library Preparation for Low DNA Input Whole Genome Bisulfite Sequencing

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DNA Methylation Protocols

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

Abstract

Aberrations of the DNA methylome contribute to onset and progression of diseases. Whole genome bisulfite sequencing (WGBS) is the only analytical method covering the complete methylome. Alternative methods requiring less DNA than WGBS analyze only a minor portion of the methylome and do not cover important regulatory features like enhancers and noncoding RNAs. In tagmentation-based WGBS (TWGBS), several DNA and time-consuming steps of the conventional WGBS library preparation are circumvented by the use of a hyperactive transposase, which simultaneously fragments DNA and appends sequencing adapters. TWGBS requires only nanogram amounts of DNA and, thus, is well suited to study precious biological specimens such as sorted cells or micro-dissected tissue samples.

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Acknowledgments

We gratefully acknowledge excellent technical support by Marion Bähr, Monika Helf and helpful discussions with Daniel Lipka. We also acknowledge the excellent support from the sequencing core facility at the DKFZ, particularly from André Götze. Work in the Plass laboratory was supported by the Helmholtz Foundation and the German Federal Ministry of Education and Science in the program for medical genome research (FKZ: 01KU1001A). Q.W. obtained support by the Humboldt Research Fellowship for Postdoctoral Researchers. A.A. is funded by an NSF Graduate Research Fellowship.

Author Contributions: D.W., Q.W., A. A. and J.S. conceived the study. S.W., R.E. and C.P. contributed materials. D.W. did the experiments and analyzed data. D.W. and C.P. wrote the manuscript.

Competing financial interests: The authors declare no competing financial interests. A provisional patent application has been deposited for aspects of these methods (A.A., J.S.).

Author information

Authors and Affiliations

  1. Division of Epigenomics and Cancer Risk Factors, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120, Heidelberg, Germany

    Dieter Weichenhan & Christoph Plass

  2. Applied Bioinformatics, German Cancer Research Center (DKFZ), Heidelberg, Germany

    Qi Wang

  3. Department of Molecular & Medical Genetics and the Knight Cardiovascular Institute, Oregon Health and Science University, Portland, OR, USA

    Andrew Adey

  4. Core Facility High Throughput Sequencing, German Cancer Research Center (DKFZ), Heidelberg, Germany

    Stephan Wolf

  5. Department of Genome Sciences, University of Washington, Seattle, WA, USA

    Jay Shendure

  6. Theoretical Bioinformatics, German Cancer Research Center (DKFZ), Heidelberg, Germany

    Roland Eils

  7. Division of Applied Bioinformatics, German Cancer Research Center (DKFZ), and German Cancer Consortium (DKTK), Heidelberg, Germany

    Qi Wang

Authors
  1. Dieter Weichenhan
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  2. Qi Wang
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  3. Andrew Adey
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  4. Stephan Wolf
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  5. Jay Shendure
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  6. Roland Eils
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  7. Christoph Plass
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Corresponding author

Correspondence to Dieter Weichenhan .

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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Weichenhan, D. et al. (2018). Tagmentation-Based Library Preparation for Low DNA Input Whole Genome Bisulfite Sequencing. 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_6

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  • DOI: https://doi.org/10.1007/978-1-4939-7481-8_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7479-5

  • Online ISBN: 978-1-4939-7481-8

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