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Biotechniques. 2013 Oct;55(4):181-97. doi: 10.2144/000114087.

Optimizing methodologies for PCR-based DNA methylation analysis.

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Laboratory of NeuroPsychiatric Genetics, Biomedical Sciences Research Group, School of Medicine, Universidad Antonio Nariño, Bogotá, Colombia; Neurosciences Research Group, School of Medicine, Universidad Nacional de Colombia, Bogotá, Colombia; Biomedical Sciences Doctoral Program, School of Medicine, Universidad Nacional de Colombia, Bogotá, Colombia.


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.

[Indexed for MEDLINE]

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