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Updated: Apr 6, 2026

DNA Methylation: Bisulphite Modification and Analysis

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

The BisPCR(2) method for targeted bisulfite sequencing.

Diana L Bernstein¹, Vasumathi Kameswaran¹, John E Le Lay¹

¹Department of Genetics and Institute for Diabetes, Obesity, and Metabolism, Perelman School of Medicine, University of Pennsylvania, 3400 Civic Center Blvd., Philadelphia, PA 19104 USA.

Epigenetics & Chromatin

|August 4, 2015

Summary

This summary is machine-generated.

We developed BisPCR(2), a cost-effective targeted bisulfite sequencing method. This technique efficiently validates DNA methylation changes at specific genetic loci, improving upon previous genome-wide studies.

Keywords:

DNA methylation Next-generation sequencing Targeted bisulfite sequencing

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Last Updated: Apr 6, 2026

DNA Methylation: Bisulphite Modification and Analysis

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

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

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

Published on: February 24, 2015

Methodology for Accurate Detection of Mitochondrial DNA Methylation

Methodology for Accurate Detection of Mitochondrial DNA Methylation

Published on: May 20, 2018

Area of Science:

Epigenetics
Genomics
Molecular Biology

Background:

DNA methylation is crucial for development and disease.
Efficient and cost-effective methods for detecting DNA methylation are needed.
Next-generation sequencing (NGS) offers high resolution but can be costly genome-wide.

Purpose of the Study:

To present a simplified, targeted bisulfite sequencing approach.
To validate differential DNA methylation at type 2 diabetes risk loci.
To demonstrate a robust, efficient, and cost-effective technique for targeted bisulfite NGS.

Main Methods:

Developed BisPCR(2), a targeted bisulfite sequencing method.
Utilized sodium bisulfite conversion and two rounds of PCR for target enrichment and sample barcoding.
Applied BisPCR(2) to human islet samples.

Main Results:

Validated differential methylation at type 2 diabetes risk loci.
Confirmed some previous findings and identified novel differentially methylated CpGs.
Achieved higher sequencing depth compared to array-based approaches.

Conclusions:

BisPCR(2) is a robust, efficient, and cost-effective technique for targeted bisulfite NGS.
The method is useful for reanalyzing and validating findings from genome-wide studies.
Enables precise quantification of CpG methylation levels at specific loci.