DNA methylation is the modification of DNA by the addition of a methyl group to the 5-position of cytosines. The process is implicated in gene regulation, genomic imprinting, embryonic development, and cell growth and differentiation. Alteration of DNA methylation may lead to diseases including cancer.
As a result, it is important to investigate patterns of DNA methylation status. Different traditional techniques have been used in methylation studies, including sequencing of bisulfite-treated DNA, methylation-sensitive restriction enzymes and Southern blotting, methylation-sensitive enzymes and PCR amplification. However, these methods are labor-intensive and time-consuming. DHPLC provides an efficient alternative for rapid and reliable methylation detection.
DHPLC analyzes the methylation-specific PCR products under partially denaturing HPLC condition or the PE products under completely denaturing HPLC condition. The DHPLC method is capable of distinguishing overall methylation profiles of differentially methylated regions of imprinted genes. The technique also allows the quantification of relative amounts of methylated and unmethylated molecules.