Targeted methylation analyses: From bisulfite treatment to quantification

J Lobo, AJM Gillis, LHJ (Leendert) Looijenga

Research output: Chapter/Conference proceedingChapterAcademic

1 Citation (Scopus)

Abstract

DNA methylation constitutes the most studied epigenetic mechanism, regulating gene expression in several physiological and pathological states. Targeted methylation polymerase chain reaction (PCR)-based analyses are among the most universal and commonly used techniques in research. They can be of use for validating methylation-based biomarkers to include in clinical practice. Optimal execution and interpretation of data is fundamental for achieving accurate and reproducible results. In this chapter we describe the backbone procedures behind targeted methylation analyses: bisulfite conversion and downstream PCR-based techniques, including real-time quantitative methylation-specific PCR (qMSP) and high-resolution melting (HRM) methylation-sensitive analyses. Specifically, we give details about the protocol, discuss the pros and cons of these methodologies, and give practical tips for achieving optimal results and for troubleshooting.

Original languageEnglish
Title of host publicationTesticular Germ Cell Tumors: Methods and Protocols
Place of Publication[S.l.)
PublisherSpringer-Verlag
Pages167-180
Number of pages167
Volume2195
ISBN (Print)9781071608593
DOIs
Publication statusPublished - 2021

Publication series

SeriesMethods in Molecular Biology
Volume2195

Bibliographical note

Export Date: 24 January 2021

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