Epigenetics. 2021 Jun;16(6):587-596. doi: 10.1080/15592294.2020.1814487. Epub 2020 Aug 30.

Comparing a new method for mapping nucleosomes in simian virus 40 chromatin to standard procedures.

Epigenetics

Barry Milavetz, Jacob Haugen, Kincaid Rowbotham

PMID: 32838633 PMCID: PMC8143259 DOI: 10.1080/15592294.2020.1814487

Abstract

The location of nucleosomes in chromatin significantly impacts many biological processes including DNA replication, repair, and gene expression. A number of techniques have been developed for mapping nucleosome locations in chromatin including MN-Seq (micrococcal nuclease digestion followed by next-generation sequencing), ATAC-Seq (Assay for Transposase-Accessible Chromatin followed by next-generation sequencing), and ChIP-Seq (chromatin immunoprecipitation and fragmentation followed by next-generation sequencing). All of these techniques have been successfully used, but each with its own limitations. Recently, New England Biolabs has marketed a new kit, the NEBNext Ultra II FS Library Prep kit, for preparing libraries for next-generation sequencing from purified genomic DNA. This kit is based on a novel proprietary DNA fragmentation procedure which appears to cleave DNA that is not bound by proteins. Because DNA is fragmented directly in the FS kit, we tested whether the kit might also be useful for mapping the location of nucleosomes in chromatin. Using simian virus 40 (SV40) chromatin isolated at different times in an infection, we have compared nucleosome mapping using the NEB FS kit (referred to as FS-Seq) to MN-Seq, ATAC-Seq, and ChIP-Seq. Mapping nucleosomes using FS-Seq generated nucleosome profiles similar to those generated by ATAC-Seq and ChIP-Seq in regulatory regions of the SV40 genome. We conclude that FS-Seq is a simple, robust, cost-effective procedure for mapping nucleosomes in SV40 chromatin that should be useful for other forms of chromatin as well. We also present evidence that FS-Seq may be useful for mapping transcription factors.

Keywords: ATAC-Seq; ChIP-Seq; Mn-Seq; Nucleosome mapping; SV40

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Publication Types

• Journal Article

Grant support

• P20 GM104360/GM/NIGMS NIH HHS/United States
• R03 AI142011/AI/NIAID NIH HHS/United States
• R15 AI094441/AI/NIAID NIH HHS/United States
• U54 GM128729/GM/NIGMS NIH HHS/United States