@inbook{1c6d9d0dc0244d7a99af1677ab0b5ab8,
title = "Whole Genome Methylation Sequencing via Enzymatic Conversion (EM-seq): Protocol, Data Processing, and Analysis",
abstract = "Whole genome bisulfite sequencing (WGBS) has been the gold standard technique for base resolution analysis of DNA methylation for the last 15 years. It has been, however, associated with technical biases, which lead to overall overestimation of global and regional methylation values, and significant artifacts in extreme cytosine-rich DNA sequence contexts. Enzymatic conversion of cytosine is the newest approach, set to replace entirely the use of the damaging bisulfite conversion of DNA. The EM-seq technique utilizes TET2, T4-BGT, and APOBEC in a two-step conversion process, where the modified cytosines are first protected by oxidation and glucosylation, followed by deamination of all unmodified cytosines to uracil. As a result, EM-seq is degradation-free and bias-free, requires low DNA input, and produces high library yields with longer reads, little batch variation, less duplication, uniform genomic coverage, accurate methylation over a larger number of captured CpGs, and no sequence-specific artifacts.",
keywords = "APOBEC, Bismark, BS-seq, DNA methylation, EM-seq, Enzymatic methylome sequencing, Nextflow, SeqMonk, T4-BGT, TET2, WGBS",
author = "Olova, \{Nelly N.\} and Simon Andrews",
note = "The authors thank Louise Williams, Max Fritsch, and Michael Skelly from New England Biolabs Inc. for valuable discussions, critical reading, and provision of materials, as well as Phil Ewels from Seqera Labs for valuable discussions and critical reading of the manuscript.",
year = "2025",
doi = "10.1007/978-1-0716-4192-7\_5",
language = "English",
isbn = "978-1-0716-4191-0",
volume = "2866",
series = "Methods in molecular biology (Clifton, N.J.)",
publisher = "Humana Press",
pages = "73--98",
editor = "Carabetta, \{Valerie J.\} and Olaitan Akintunde",
booktitle = "High Throughput Gene Screening",
}