How Does MS-HRM Work?

How Does MS-HRM Work?

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  2. How does MS-HRM work?

Methylation-Sensitive High-Resolution Melting (MS-HRM) is based on a PCR methylation detection method.


The number of cytosines (C) in a PCR product, amplified from a sodium bisulfite modified template, depends on the methylation status of cytosines in the un-modified template.

Sodium bisulfite preserves the methylated cytosines and modifies non-methylated cytosines into uracil, which subsequently is substituted with thymine during PCR.

Thus, if the locus is methylated, a C rich PCR product is obtained after PCR amplification (see A below), and a T rich PCR product if the locus was non-methylated (see B below).

These PCR products will display different melting profiles (see also: What is DNA melting?) when subjected to Methylation-Sensitive High-Resolution Melting (MS-HRM) analyses.

The PCR product amplified from the non-methylated version of a specific locus will have relatively low melting temperature and melts earlier in the temperature gradient (Figure 1 – red curve) than the PCR product amplified from the methylated version of the same locus, which will melt at relatively higher temperature (Figure 1 – blue curve).

In summary, MS-HRM is a PCR methylation detection method, dependant on bisulfite modification, and enables cost effective, fast, and highly sensitive assessment of locus specific methylation status.

A: PCR product obtained from the methylated version of the locus:


B: PCR product obtained from the non-methylated version of the locus:


Figure 1:

Further reading:

  1. MS-HRM protocol in Nature Protocols.


  1. MS-HRM in Nucleic Acid Research


  1. MS-HRM protocol in Methods in Molecular Biology



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