DNA methylation of cytosine by methyltransferase is one of the epigenetic changes that occur in CpG repeat sites. DNA methylation changes occur during carcinogenesis, and this mechanism has revealed several methylation biomarkers.

Bisulfite conversion is the most commonly used laboratory method for DNA methylation detection. If a site is methylated at a cytosine, the cytosine is converted to uracil after bisulfite conversion. If the site is not methylated, it remains as cytosine. Therefore, bisulfite conversion can reveal sequence changes and determine the methylation level.

MS-HRM provides an efficient and cost-effective measurement of the methylation level of a sample. It uses a primer called a methylation-independent primer (MIP) that can amplify the target sequence, regardless of the methylation status of the target. The PCR product created by the MIP undergoes a slow increase in temperature, and these changes in the melting temperature of the product indicate the degree of methylation.