Direct DNA methylation sequencing

Overview:

Direct DNA Methylation Sequencing using Oxford Nanopore technology enables measurement of methylation marks without bisulfite conversion or PCR, preserving native DNA integrity. Nanopore sensors detect methylation states directly from electrical signal shifts, providing long-read methylation profiles across kilobase- to megabase-long molecules. High molecular-weight DNA is essential. Coverage requirements vary by application and can range from low-coverage (5–10x) for global methylation estimation to higher depths for locus-specific quantification. This method supports phasing of genetic and epigenetic variants, long-range methylation haplotypes, detection of methylation across repetitive and structurally complex regions, and integrative analysis of methylation with structural variation. Applications include cancer epigenomics, imprinting and allele-specific methylation studies, developmental epigenetics, microbial epigenome profiling, and rapid methylation analysis in real-time sequencing workflows.

Workflow:

DNA extraction, library preparation for ONT, sequencing with real-time base modification calling.