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Nanopore Sequencing and Data Analysis for Base-Resolution Genome-Wide 5-Methylcytosine Profiling


Whole-genome bisulfite sequencing (WGBS) is currently the gold standard for DNA methylation (5-methylcytosine, 5mC) profiling; however, the destructive nature of sodium bisulfite results in DNA fragmentation and subsequent biases in sequencing data. Such issues have led to the development of bisulfite-free methods for 5mC detection. Nanopore sequencing is a long read nondestructive approach that directly analyzes DNA and RNA fragments in real time. Recently, computational tools have been developed that enable base-resolution detection of 5mC from Oxford Nanopore sequencing data. In this chapter, we provide a detailed protocol for preparation, sequencing, read assembly, and analysis of genome-wide 5mC using Nanopore sequencing technologies.

Type Journal
ISBN 1940-6029 (Electronic) 1064-3745 (Linking)
Authors Angeloni, A.; Ferguson, J.; Bogdanovic, O.
Publisher Name Methods in Molecular Biology
Published Date 2022-02-28
Published Volume 2458
Published Pages 75-94
Status Published in-print
DOI 10.1007/978-1-0716-2140-0_5
URL link to publisher's version