high resolution melting analysis

high resolution melting analysis (HRM):
-used to detect SNP
-must use a saturating dye (e.g. Evagreen)
-after real-time PCR, normalize a melt curve, suppress the curve that differs the most.

Methylation Sensitive (MS)-HRM:

-Bisulfite treatment of DNA converts unmethylated Cytosine to Uracil.
— Methylated Cytosine are protected and remain intact.

-After PCR amplification all Uracils are converted Thymine.

-perform DNA sequence analysis, if T, that means the DNA is unmethylated.
if C, the DNA is methylated.