Quantitative DNA Methylation Analysis by Pyrosequencing

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...

Full description

Saved in:
Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 1315; p. 175
Main Authors Roessler, Jessica, Lehmann, Ulrich
Format Journal Article
LanguageEnglish
Published United States 2015
Subjects
CpG Islands - genetics
DNA - genetics
DNA - isolation & purification
DNA Methylation - drug effects
Polymerase Chain Reaction
Sequence Analysis, DNA - methods
Sulfites - pharmacology
Online AccessGet more information

Cover

More Information
Summary: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.
ISSN:1940-6029
DOI:10.1007/978-1-4939-2715-9_13