Related Experiment Video
Updated: Mar 13, 2026
13:47
Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution
Published on: February 24, 2015
26.5K
The Combined Bisulfite Restriction Analysis (COBRA) Assay for the Analysis of Locus-Specific Changes in Methylation
Andriy Bilichak1, Igor Kovalchuk2
1Agriculture and Agri-Food Canada, Lethbridge Research Centre, Lethbridge, AB, Canada. andrii.bilichak@agr.gc.ca.
Methods in Molecular Biology (Clifton, N.J.)
|October 23, 2016
Summary
This study introduces the combined bisulfite restriction analysis (COBRA) assay for precise DNA methylation analysis. COBRA enables researchers to determine methylation status at specific genomic loci, aiding gene expression studies.
Area of Science:
- Epigenetics
- Molecular Biology
- Genomics
Background:
- DNA methylation is a key epigenetic regulator of gene expression.
- Methylation patterns influence chromatin structure, gene activity, and genome stability.
- Analyzing locus-specific methylation is crucial for understanding these effects.
Purpose of the Study:
- To describe the combined bisulfite restriction analysis (COBRA) assay.
- To provide a detailed protocol for analyzing DNA methylation at specific genomic loci.
Main Methods:
- Bisulfite conversion of DNA to alter non-methylated cytosines.
- Locus-specific PCR amplification of converted DNA.
- Restriction digestion, gel electrophoresis, and quantitative analysis of DNA methylation patterns.
Main Results:
- The COBRA assay allows for the analysis of DNA methylation status at specific loci.
- The protocol involves bisulfite conversion, PCR, restriction digestion, and quantification.
- Quantitative analysis can be performed using software like ImageJ.
Conclusions:
- The COBRA assay is a valuable tool for assessing locus-specific DNA methylation.
- This method facilitates research into the functional roles of DNA methylation in gene regulation and genome stability.
