Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands

摘要: Abstract Precise mapping of DNA methylation patterns in CpG islands has become essential for understanding diverse biological processes such as the regulation imprinted genes, X chromosome inactivation, and tumor suppressor gene silencing human cancer. We describe a new method, MSP (methylation-specific PCR), which can rapidly assess status virtually any group sites within island, independent use methylation-sensitive restriction enzymes. This assay entails initial modification by sodium bisulfite, converting all unmethylated, but not methylated, cytosines to uracil, subsequent amplification with primers specific methylated versus unmethylated DNA. requires only small quantities DNA, is sensitive 0.1% alleles given island locus, be performed on extracted from paraffin-embedded samples. eliminates false positive results inherent previous PCR-based approaches relied differential enzyme cleavage distinguish In this study, we demonstrate identify promoter region hypermethylation changes associated transcriptional inactivation four important genes (p16, p15, E-cadherin, von Hippel-Lindau)