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Related Concept Videos

High-Resolution Mass Spectrometry (HRMS)01:15

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The resolution of a mass spectrometer depends on the efficiency of separating ions with different ion masses. The mass of an atom is approximated to the sum of the masses of protons and neutrons inside, considering the masses of protons and neutrons as equal. However, the masses of the proton (1.6726 × 10−24 g) and neutron (1.6749 × 10−24 g) are not truly equal. There is a minor error in the expression of atomic masses relative to the simplest atom of hydrogen. For...
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Related Experiment Video

Updated: Feb 17, 2026

Correlating Gene-specific DNA Methylation Changes with Expression and Transcriptional Activity of Astrocytic KCNJ10 Kir4.1
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Methylation-Sensitive High Resolution Melting (MS-HRM).

Dianna Hussmann1, Lise Lotte Hansen2

  • 1Institute of Biomedicine, Aarhus University, Bartholins Allé 6, Aarhus C, 8000, Denmark.

Methods in Molecular Biology (Clifton, N.J.)
|December 11, 2017
PubMed
Summary

Methylation-Sensitive High Resolution Melting (MS-HRM) is a sensitive PCR method for detecting DNA methylation. This technique can identify low levels of methylated DNA, aiding in disease diagnosis and environmental exposure studies.

Keywords:
DNA methylationHeterogeneous methylationMS-HRMPCR biasPrimer design

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Area of Science:

  • Epigenetics
  • Molecular Biology
  • Biotechnology

Background:

  • DNA methylation is a crucial epigenetic modification involved in gene regulation and disease.
  • Accurate detection of methylation levels is essential for understanding biological processes and diagnosing diseases.
  • Existing methods for methylation analysis can be complex, time-consuming, or lack sensitivity.

Purpose of the Study:

  • To introduce and describe Methylation-Sensitive High Resolution Melting (MS-HRM) as a sensitive, in-tube PCR-based method for detecting DNA methylation.
  • To highlight the unique primer design and bisulfite treatment steps that enhance assay sensitivity.
  • To showcase the broad applicability of MS-HRM in various diagnostic and research contexts.

Main Methods:

  • Utilizes Methylation-Sensitive High Resolution Melting (MS-HRM), an in-tube, PCR-based assay.
  • Employs a unique primer design that anneals to both methylated and unmethylated alleles, increasing sensitivity.
  • Incorporates bisulfite treatment of DNA to differentiate between methylated and unmethylated sequences, enabling separation by high-resolution melting.

Main Results:

  • Achieves high sensitivity, detecting as little as 0.1-1% methylated alleles in an unmethylated background.
  • Demonstrates utility in detecting cancer biomarkers noninvasively in urine, stool, and buccal mucosa samples.
  • Proves effective for rapid diagnosis of imprinted diseases and validation of epigenome-wide studies.

Conclusions:

  • MS-HRM is a highly sensitive and fast method for detecting DNA methylation at specific loci.
  • Its ability to detect low levels of methylated DNA makes it valuable for noninvasive disease biomarker discovery.
  • MS-HRM is a key tool for investigating links between environmental factors, epigenetics, and disease development.