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Related Concept Videos

Phase II Reactions: Methylation Reactions01:17

Phase II Reactions: Methylation Reactions

Methylation is a phase II biotransformation process involving the attachment of a methyl group to a substrate. Enzymes known as methyltransferases orchestrate this reaction.
The mechanism of methylation unfolds in two stages. The first stage sees a methyltransferase enzyme facilitating the transfer of a methyl group from S-adenosylmethionine (SAM) to the substrate, forming S-adenosylhomocysteine (SAH). The second stage involves further metabolism of SAH into homocysteine, which can be recycled...

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Related Experiment Video

Updated: May 28, 2026

DNA Methylation: Bisulphite Modification and Analysis
12:34

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

DNA methylation: bisulphite modification and analysis.

Kate Patterson1, Laura Molloy, Wenjia Qu

  • 1Epigenetics Group, Cancer Research Program, Garvan Institute of Medical Research.

Journal of Visualized Experiments : Jove
|November 2, 2011
PubMed
Summary

Bisulphite conversion is the gold standard for detecting DNA methylation (5-methylcytosine). This method precisely quantifies methylation at single nucleotide resolution, crucial for understanding its role in development and disease.

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

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Last Updated: May 28, 2026

DNA Methylation: Bisulphite Modification and Analysis
12:34

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

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

Targeted DNA Methylation Analysis by Next-generation Sequencing

Published on: February 24, 2015

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution
13:47

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

Published on: February 24, 2015

Area of Science:

  • Epigenetics and Molecular Biology
  • Genomics and Bioinformatics

Background:

  • Epigenetics involves heritable gene function changes independent of DNA sequence.
  • DNA methylation, primarily cytosine methylation (5-MeC), regulates gene expression.
  • CpG dinucleotides are key targets for DNA methylation, concentrated in promoters and repetitive regions.

Purpose of the Study:

  • To detail the gold standard bisulphite conversion protocol for DNA methylation analysis.
  • To enable precise, efficient, and reproducible detection and quantification of 5-methylcytosine (5-MeC).
  • To facilitate understanding of DNA methylation's biological roles and its disruption in diseases like cancer.

Main Methods:

  • Bisulphite conversion of genomic DNA.
  • Chemical process involves cytosine deamination to uracil by sodium bisulphite, while 5-MeC remains unchanged.
  • Post-PCR analysis, including cloning and sequencing, for single nucleotide resolution of methylation patterns.

Main Results:

  • Bisulphite conversion allows differentiation of methylated (cytosine) and unmethylated (thymine) CpG sites after PCR.
  • This method provides single nucleotide resolution for DNA methylation analysis.
  • Established protocol widely used for generating new DNA methylation data.

Conclusions:

  • Bisulphite conversion is essential for accurate DNA methylation analysis.
  • The protocol's ability to resolve methylation at single nucleotide resolution is critical for research.
  • This technique is fundamental for studying epigenetic regulation in normal development and disease states.