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Related Experiment Video

Updated: May 19, 2026

DNA Methylation: Bisulphite Modification and Analysis
12:34

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

DNA methylation analysis triggered by bulge specific immuno-recognition.

Ryoji Kurita1, Osamu Niwa

  • 1National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 6, Tsukuba, Ibaraki, Japan. r.kurita@aist.go.jp

Analytical Chemistry
|August 14, 2012
PubMed
Summary

This study introduces a novel method for detecting DNA cytosine methylation status using an anti-methylcytosine antibody and a DNA bulge. This technique accurately identifies methylation at specific DNA sequences without bisulfite treatment.

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Area of Science:

  • Molecular Biology
  • Epigenetics
  • Biotechnology

Background:

  • Cytosine methylation is a crucial epigenetic modification impacting gene expression.
  • Accurate detection of DNA methylation status is vital for understanding various biological processes and diseases.
  • Current methods often require harsh treatments like bisulfite conversion or are limited in sequence specificity.

Purpose of the Study:

  • To develop a novel, sequence-selective method for discriminating DNA cytosine methylation status.
  • To utilize an anti-methylcytosine antibody and a DNA bulge strategy for enhanced specificity.
  • To enable methylation detection without conventional bisulfite treatment, PCR, or electrophoresis.

Main Methods:

  • Hybridization of DNA with a bulge-inducing sequence to position target methylcytosine in a bulge.

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Comprehensive DNA Methylation Analysis Using a Methyl-CpG-binding Domain Capture-based Method in Chronic Lymphocytic Leukemia Patients
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Comprehensive DNA Methylation Analysis Using a Methyl-CpG-binding Domain Capture-based Method in Chronic Lymphocytic Leukemia Patients

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Last Updated: May 19, 2026

DNA Methylation: Bisulphite Modification and Analysis
12:34

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

Immunostaining for DNA Modifications: Computational Analysis of Confocal Images
09:42

Immunostaining for DNA Modifications: Computational Analysis of Confocal Images

Published on: September 7, 2017

Comprehensive DNA Methylation Analysis Using a Methyl-CpG-binding Domain Capture-based Method in Chronic Lymphocytic Leukemia Patients
13:21

Comprehensive DNA Methylation Analysis Using a Methyl-CpG-binding Domain Capture-based Method in Chronic Lymphocytic Leukemia Patients

Published on: June 16, 2017

  • Affinity measurement of anti-methylcytosine antibody binding to methylcytosine in the bulge versus duplex regions.
  • Combination of the bulge-specific assay with absorbance measurement using horseradish peroxidase-labeled secondary antibody.

    • Main Results:

    • The antibody showed significantly higher affinity for methylcytosine in a DNA bulge compared to a double-stranded DNA duplex.
    • The method achieved sequence-selective discrimination of cytosine methylation status at a single base level.
    • The assay demonstrated robustness against various types of interfering genomic DNA contamination.

    Conclusions:

    • The developed bulge-specific assay provides a highly selective and sensitive method for detecting DNA cytosine methylation.
    • This technique offers a promising alternative to conventional methods, simplifying the detection process.
    • The approach has potential applications in epigenetic research and diagnostics, particularly for promoter regions like MGMT.