@inbook{b1c36cc18d264681aa90934cce3851ec,
title = "Targeted Methylation Analyses: From Bisulfite Treatment to Quantification",
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.",
keywords = "DNA Methylation, Epigenesis, Genetic, Epigenomics/methods, Humans, Polymerase Chain Reaction, Real-Time Polymerase Chain Reaction, Sequence Analysis, DNA",
author = "Jo{\~a}o Lobo and Gillis, {Ad J.M.} and Looijenga, {Leendert H.J.}",
note = "Publisher Copyright: {\textcopyright} Springer Science+Business Media, LLC, part of Springer Nature 2021.",
year = "2021",
doi = "10.1007/978-1-0716-0860-9_12",
language = "English",
volume = "2195",
series = "Methods in molecular biology (Clifton, N.J.)",
publisher = "Humana Press Inc.",
pages = "167--180",
booktitle = "Methods in Molecular Biology",
}