Quantitation of DNA methylation by melt curve analysis.

ABSTRACT:

Background

Methylation of DNA is a common mechanism for silencing genes, and aberrant methylation is increasingly being implicated in many diseases such as cancer. There is a need for robust, inexpensive methods to quantitate methylation across a region containing a number of CpGs. We describe and validate a rapid, in-tube method to quantitate DNA methylation using the melt data obtained following amplification of bisulfite modified DNA in a real-time thermocycler.

Methods

We first describe a mathematical method to normalise the raw fluorescence data generated by heating the amplified bisulfite modified DNA. From this normalised data the temperatures at which melting begins and finishes can be calculated, which reflect the less and more methylated template molecules present

SUBMITTER: Smith E 

PROVIDER: S-EPMC2679043 | biostudies-literature | 2009 Apr

REPOSITORIES: biostudies-literature