Optimizing methodologies for PCR-based DNA methylation analysis

Comprehensive analysis of DNA methylation patterns is critical for understanding the molecular basis of many human diseases. While hundreds of PCR-based DNA methylation studies are published every year, the selection and implementation of appropriate methods for these studies can be challenging for...

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Bibliographic Details
Published inBioTechniques Vol. 55; no. 4; pp. 181 - 197
Main Authors Hernández, Hernán G, Tse, M. Yat, Pang, Stephen C, Arboleda, Humberto, Forero, Diego A
Format Journal Article
LanguageEnglish
Published England Future Science Ltd 01.10.2013
Taylor & Francis Group
Subjects
5-methylcytosine
DNA methylation
DNA Methylation - genetics
epigenomics
High-Throughput Nucleotide Sequencing - methods
Humans
MethyLight
MS-HRM
Nucleic Acid Denaturation - genetics
polymerase chain reaction
Polymerase Chain Reaction - methods
reference standards
Sulfites - chemistry
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Summary:Comprehensive analysis of DNA methylation patterns is critical for understanding the molecular basis of many human diseases. While hundreds of PCR-based DNA methylation studies are published every year, the selection and implementation of appropriate methods for these studies can be challenging for molecular genetics researchers not yet familiar with methylation analysis. Here we review the most commonly used PCR-based DNA methylation analysis techniques: bisulfite sequencing PCR (BSP), methylation specific PCR (MSP), MethyLight, and methylation-sensitive high resolution melting (MS-HRM). We provide critical analysis of the strengths and weaknesses of each approach as well as a series of guidelines to assist in selecting and implementing an appropriate method.
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ISSN:0736-6205
1940-9818
DOI:10.2144/000114087