Library Preparation for Genome-Wide DNA Methylation Profiling
Journal
Bio-protocol
Journal Volume
15
Journal Issue
1383
Start Page
e5488
ISSN
2331-8325
Date Issued
2025
Author(s)
Abstract
DNA methylation is a fundamental epigenetic mark with critical roles in epigenetic regulation, development, and genome stability across diverse organisms. Whole genome bisulfite sequencing (WGBS) enables single-base resolution mapping of cytosine methylation patterns and has become a standard method in epigenomics. This protocol provides a detailed, step-by-step workflow for WGBS library construction starting from genomic DNA. It includes steps of RNaseA treatment, DNA shearing, end-repair and A-tailing, adapter ligation, bisulfite conversion, library amplification, and quantification. Notably, the method uses self-prepared reagents and customizable index systems, avoiding the constraints of commercial library preparation kits. This flexibility supports cost-effective, scalable methylome profiling, suitable for diverse experimental designs, including high-throughput multiplexed sequencing.
Subjects
DNA methylation
Epigenetics
Epigenomics
Library preparation
Next generation sequencing
Whole genome bisulfite sequencing
Publisher
Bio-Protocol, LLC
Description
Article number e5488
Type
journal article