Introduction
Methylation is a normally occurring modification to DNA. It is characterized by the biochemical addition of a methyl group to the cytosine 5-carbon in cytosine-phosphate-guanosine (CpG) dinucleotides via a methytransferase enzyme. DNA methylation plays an important role in regulating gene expression. Aberrant DNA methylation has been implicated in many disease processes, including cancer, obesity, and addiction. It’s also a common subject of agrigenomic investigations into responses to drought, temperature extremes, and other environmental changes.
High-throughput technologies such as next-generation sequencing (NGS) and microarrays enable researchers to perform genome-wide methylation profiling, and researchers can also use high-performance capillary electrophoresis and methylation-sensitive arbitrarily primed PCR to quantify DNA methylation.
For gene-specific methylation analysis, the most popular method is the bisulfite reaction-based method, this method use sodium bisulfite to convert the unmethylated cytosines on DNA to uracils, thus the modified DNA template is distinguishable from the original template. The methylated cytosines could be detected using methylation specific PCR (MSP). Further PCR amplification and sequencing allow for this differential conversion to be detected.
Fig. 2 Determination of differential methylation with bisulfite sequencing.
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