The OneStep PLUS qMethyl PCR Kit provides a simple, straightforward, and bisulfite-free procedure for the quantification of DNA methylation at custom-selected genomic regions. This bisulfite-free assessment is accomplished by splitting the DNA to be tested into two parts: a “Test Reaction” and a “Reference Reaction”. DNA in the Test Reaction is digested with Methylation Sensitive Restriction Enzymes (MSREs) while DNA in the Reference Reaction is not. The DNA from both reactions is then amplified using Real-Time PCR. The difference in Cycle Threshold (Ct) values between the Reference and Test reactions is used to calculate the percentage of DNA methylation at the selected genomic region.