Targeted Methylation Analyses: From Bisulfite Treatment to Quantification

João Lobo, Ad J.M. Gillis, Leendert H.J. Looijenga

Onderzoeksoutput: Hoofdstuk in Boek/Rapport/CongresprocedureHoofdstukpeer review

1 Citaat (Scopus)


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.

Originele taal-2Engels
TitelMethods in Molecular Biology
UitgeverijHumana Press Inc.
Aantal pagina's14
StatusGepubliceerd - 2021

Publicatie series

NaamMethods in molecular biology (Clifton, N.J.)
UitgeverijHumana Press
ISSN van geprinte versie1064-3745


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