Whole-genome bisulfite sequencing

Overview:

Whole-Genome Bisulfite Sequencing provides a base-resolution map of DNA methylation across the entire genome. Through sodium bisulfite conversion, unmethylated cytosines are converted to uracil, enabling quantification of methylated and unmethylated CpG, CHG, and CHH sites. Sequencing is generally performed using Illumina short-read platforms due to their high accuracy, with typical coverage ranging from 20× to 60× depending on experimental goals. High-quality genomic DNA is required, as bisulfite treatment can significantly fragment DNA and reduce complexity. Libraries can be prepared using PCR-free or PCR-based kits depending on input availability; typical inputs range from 100 ng to >1 µg of DNA. WGBS enables global methylation profiling in mammalian cells, plants, and microbial genomes, supporting studies in epigenomic regulation, developmental biology, environmental epigenetics, cancer epigenomics, and biomarker discovery. It allows detection of differentially methylated regions (DMRs), identification of epigenetically regulated promoters and enhancers, and methylation mapping in intergenic or repetitive regions, among other applications.

Workflow:

Genomic DNA extraction, bisulfite conversion, library preparation, sequencing.