Question

What is a methylation-specific PCR?

Answer 1

Methylation-specific PCR:

1. It is a type of PCR (polymerase chain reaction) used for the analysis of DNA methylation patterns in CpG-rich regions of the promoter.
2. In this method, one methylated and unmethylated primer is used.
3. The unmethylated primer of PCR amplifies the sodium bisulfite converted DNA that is present in unmethylated DNA and the methylated primer amplifies the sodium bisulfite converted DNA that is present in methylated DNA.
4. To differentiate between methylated and unmethylated DNA, the primers should contain three CpG nucleotides in the 3' end of the primer and should contain a large number of non-CpG cytosines in the template.
5. For achieving good results the length of both primers should be at least 23-24 base pairs.