text: A further method to differentiate converted from unconverted bisulfite-treated DNA is using high-resolution melting analysis (HRM), a quantitative PCR-based technique initially designed to distinguish SNPs.  The PCR amplicons are analyzed directly by temperature ramping and resulting liberation of an intercalating fluorescent dye during melting.  The degree of methylation, as represented by the C-to-T content in the amplicon, determines the rapidity of melting and consequent release of the dye.  This method allows direct quantitation in a single-tube assay, but assesses methylation in the amplified region as a whole rather than at specific CpG sites.