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Quantitative DNA Methylation Analysis by Pyrosequencing®

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Pyrosequencing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1315))

Abstract

DNA methylation is an epigenetic mark playing an important role in development and disease. Aberrant DNA methylation was identified as an alternative mechanism for gene inactivation complementing deletions and mutations in cancer initiation and progression. However, to accurately compare differences in DNA methylation among various tissue types, adequate quantitative approaches are required. Pyrosequencing®, as a sequencing-by-synthesis method, allows such quantification with single CpG resolution and the ability for threshold determination. This book chapter provides a detailed protocol for DNA methylation analysis by Pyrosequencing, including information on assay design and practical procedure. Additionally, emphasis is placed on the discussion of strengths and weaknesses of the methodology.

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Correspondence to Ulrich Lehmann .

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© 2015 Springer Science+Business Media New York

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Roessler, J., Lehmann, U. (2015). Quantitative DNA Methylation Analysis by Pyrosequencing® . In: Lehmann, U., Tost, J. (eds) Pyrosequencing. Methods in Molecular Biology, vol 1315. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2715-9_13

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  • DOI: https://doi.org/10.1007/978-1-4939-2715-9_13

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2714-2

  • Online ISBN: 978-1-4939-2715-9

  • eBook Packages: Springer Protocols

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