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Updated: May 7, 2026

Targeted DNA Methylation Analysis by Next-generation Sequencing

Targeted DNA Methylation Analysis by Next-generation Sequencing

Published on: February 24, 2015

Tagmentation-based whole-genome bisulfite sequencing.

Qi Wang¹, Lei Gu, Andrew Adey

¹Theoretical Bioinformatics, German Cancer Research Center (DKFZ), Heidelberg, Germany.

Nature Protocols

|September 28, 2013

Summary

This summary is machine-generated.

Tagmentation-based whole-genome bisulfite sequencing (T-WGBS) enables comprehensive methylome analysis using minimal DNA. This cost-effective method accelerates the study of DNA methylation patterns in precious samples.

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DNA Methylation: Bisulphite Modification and Analysis

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Last Updated: May 7, 2026

Targeted DNA Methylation Analysis by Next-generation Sequencing

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DNA Methylation: Bisulphite Modification and Analysis

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

Area of Science:

Epigenetics
Genomics
Molecular Biology

Background:

DNA methylation, specifically 5-methylcytosine in CpG sites, is crucial in human diseases.
Understanding the complete methylome requires quantitative analysis of all CpG sites.
Conventional whole-genome bisulfite sequencing (WGBS) is limited by high DNA input, cost, and resource requirements.

Purpose of the Study:

To introduce and validate a novel tagmentation-based whole-genome bisulfite sequencing (T-WGBS) protocol.
To enable comprehensive methylome analysis from limited DNA quantities.
To provide a more accessible method for studying genome-wide DNA methylation.

Main Methods:

Development of a tagmentation-based WGBS (T-WGBS) protocol using a hyperactive Tn5 transposase.
Single-step DNA fragmentation and adapter ligation using Tn5 transposase.
Comparison of T-WGBS with conventional WGBS to assess reliability.

Main Results:

T-WGBS requires significantly less input DNA (≤20 ng) compared to conventional WGBS.
The T-WGBS protocol is reliable and comparable to standard WGBS.
Library preparation for T-WGBS can be completed in 2 days.

Conclusions:

T-WGBS offers a robust and efficient method for comprehensive methylome analysis.
This protocol significantly reduces DNA input requirements, making methylome studies feasible for precious samples.
T-WGBS lowers barriers to genome-wide DNA methylation analysis, aiding disease research.