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Related Experiment Video

Updated: Jun 26, 2026

DNA Methylation: Bisulphite Modification and Analysis
12:34

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

Methylation-sensitive high-resolution melting.

Tomasz K Wojdacz1, Alexander Dobrovic, Lise Lotte Hansen

  • 1Institute of Human Genetics, University of Aarhus, The Bartholin Building, Wilhelm Meyers Allé, Bygn. 1242, DK-8000 Aarhus C, Denmark. wojdacz@humgen.au.dk

Nature Protocols
|January 31, 2009
PubMed
Summary

Methylation-sensitive high-resolution melting (MS-HRM) detects DNA methylation by analyzing PCR product melting profiles. This cost-efficient method accurately determines methylation status in under 3 hours.

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Last Updated: Jun 26, 2026

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Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

Published on: February 24, 2015

Area of Science:

  • Molecular Biology
  • Epigenetics
  • Biochemistry

Background:

  • DNA methylation patterns are crucial epigenetic modifications influencing gene expression.
  • Traditional methods for DNA methylation analysis are often labor-intensive and time-consuming.
  • Distinguishing between methylated and unmethylated DNA is essential for understanding various biological processes.

Purpose of the Study:

  • To introduce and validate a novel, rapid, and cost-efficient protocol for DNA methylation analysis.
  • To enable sensitive, single-locus determination of DNA methylation status.
  • To provide a labor-efficient alternative to existing methylation detection methods.

Main Methods:

  • Sodium bisulfite modification of template DNA.
  • PCR amplification using primers designed for both methylated and unmethylated templates.
  • High-resolution melting (HRM) analysis of PCR products to compare melting profiles.

Main Results:

  • Methylation-dependent differences in PCR product base composition lead to distinct melting profiles.
  • The MS-HRM protocol allows for in-tube determination of methylation status.
  • The method demonstrates high sensitivity and efficiency for single-locus methylation studies.

Conclusions:

  • MS-HRM is a highly sensitive, labor- and cost-efficient technique for determining DNA methylation status.
  • The protocol offers a rapid (under 3 hours) and reliable method for epigenetic analysis.
  • MS-HRM provides a valuable tool for single-locus methylation studies based on DNA high-resolution melting technology.