title: PCR Techniques in Characterizing DNA Methylation.
abstract: DNA methylation was the first epigenetic mark to be discovered, involving the addition of a methyl group to the 5' position of cytosine by DNA methyltransferases, and can be inherited through cell division. DNA methylation plays an important role in normal human development and is associated with the regulation of gene expression, tumorigenesis, and other genetic and epigenetic diseases. Differential methylation is now known to play a central role in the development and outcome of most if not all human malignancies.Bisulfite conversion is a commonly used approach for gene-specific DNA methylation analysis. Treatment of DNA with bisulfite converts cytosine to uracil while leaving 5-methylcytosine intact, allowing for single-nucleotide resolution information about the methylated areas of DNA. PCR-based methods are routinely used to study DNA methylation on a gene-specific basis, after bisulfite treatment. Variations of this method include bisulfite sequencing, methylation-specific PCR, real-time PCR-based MethyLight, and methylation-sensitive high-resolution melting PCR. Several whole-epigenome profiling technologies such as MethylC-seq reduced representation bisulfite sequencing (RRBS) and the Infinium Human methylation 450 K bead chip are now available allowing for the identification of epigenetic drivers of disease processes as well as biomarkers that could potentially be integrated into clinical practice.
authors: Department of Translational Molecular Pathology, The University of Texas MD Anderson Cancer Centre, 2130 W. Holcombe Blvd, Life Science Plaza 11.5008, Houston, TX, 77030, USA. kwani@mdanderson.org.
authors: Khalida
authors: K
authors: Wani
authors: Princess Margaret Cancer Center and Ontario Cancer Institute, 610 University Avenue, Toronto, ON, Canada, M5G 2M9.
authors: Kenneth D
authors: KD
authors: Aldape
mesh_terms: D018899:CpG Islands; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D057890:Epigenomics / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods
keywords: Bisulfite conversion; Bisulfite sequencing; CpG islands; DNA methylation; MethyLight; Methylation sensitive-high-resolution melting (MS-HRM); Methylation-specific PCR (MSP); Real-time MSP; Whole-genome methylation profiling