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Related Experiment Video

Updated: Dec 10, 2025

DNA Methylation: Bisulphite Modification and Analysis
12:34

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

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Targeted Methylation Analyses: From Bisulfite Treatment to Quantification.

João Lobo1,2,3,4, Ad J M Gillis1, Leendert H J Looijenga5,6

  • 1Princess Máxima Center for Pediatric Oncology, Utrecht, The Netherlands.

Methods in Molecular Biology (Clifton, N.J.)
|August 28, 2020
PubMed
Summary

This chapter details targeted DNA methylation analysis methods, including bisulfite conversion and PCR-based techniques like qMSP and HRM. It provides protocols and tips for accurate biomarker validation and reproducible epigenetic research.

Keywords:
Bisulfite treatmentHigh-resolution meltingMethylationPCRStandard operating proceduresTroubleshooting

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Last Updated: Dec 10, 2025

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Published on: October 21, 2011

106.0K
Targeted DNA Methylation Analysis by Next-generation Sequencing
08:38

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Published on: February 24, 2015

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

  • Epigenetics
  • Molecular Biology
  • Biochemistry

Background:

  • DNA methylation is a key epigenetic mechanism regulating gene expression in health and disease.
  • Targeted methylation polymerase chain reaction (PCR)-based analyses are widely used in research for biomarker validation.
  • Accurate execution and interpretation are crucial for reliable epigenetic data.

Purpose of the Study:

  • To describe fundamental procedures for targeted DNA methylation analysis.
  • To provide details on bisulfite conversion and downstream PCR-based techniques.
  • To offer practical guidance for optimal results and troubleshooting in methylation analysis.

Main Methods:

  • Bisulfite conversion of DNA.
  • Real-time quantitative methylation-specific PCR (qMSP).
  • High-resolution melting (HRM) methylation-sensitive analyses.

Main Results:

  • Detailed protocols for targeted methylation analyses are presented.
  • Pros and cons of qMSP and HRM methodologies are discussed.
  • Practical tips for optimizing results and troubleshooting are provided.

Conclusions:

  • Mastery of targeted methylation analysis techniques is essential for accurate epigenetic research.
  • This chapter serves as a guide for researchers and clinicians validating methylation biomarkers.
  • Understanding these methods enhances the reliability of epigenetic data in various applications.