Direct sequencing of cytosine modification states from nanopore long-read data

Abstract

Recent developments in Oxford Nanopore sequencing include the direct and simultaneous identification of both 5-methylcytosine (5mC) and its oxidation-derivative 5-hydroxymethylcytosine (5hmC) as part of standard sequencing practices. In this study, we benchmarked nanopore modified base detection at a whole mammalian genome scale against orthologous bisulphite-based whole-genome sequencing. To complement comparisons, we demonstrate the direct and PCR-free nanopore sequencing of a 5hmC-DNA immunoprecipitation (hMeDIP) library to generate a 5hmC-enriched sequence dataset. We find highly accurate modified base detection consistent with orthologous techniques at all levels of comparison, and investigate patterns of modification symmetry and asymmetry at the level of individual DNA molecules using recent high-duplex PromethION Flow Cells.

Biography

Dominic is a third-year PhD student at the University of Bath, UK, working in the Life Science Department’s Murrell Lab. His research project focuses on the epigenetic changes of DNA involved in mammalian ageing and age-related disease.