Rapid and inexpensive preparation of genome-wide nucleosome footprints from model and non-model organisms

STAR Protoc. 2021 May 18;2(2):100486. doi: 10.1016/j.xpro.2021.100486. eCollection 2021 Jun 18.

Abstract

MNase-seq (micrococcal nuclease sequencing) is used to map nucleosome positions in eukaryotic genomes to study the relationship between chromatin structure and DNA-dependent processes. Current protocols require at least two days to isolate nucleosome-protected DNA fragments. We have developed a streamlined protocol for S. cerevisiae and other fungi which takes only three hours. Modified protocols were developed for wild fungi and mammalian cells. This method for rapidly producing sequencing-ready nucleosome footprints from several organisms makes MNase-seq faster and easier, with less chemical waste.

Keywords: Genomics; Model Organisms; Molecular Biology; Sequencing.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cell Culture Techniques / methods
  • Cells, Cultured
  • DNA / chemistry
  • DNA / genetics
  • DNA / metabolism
  • DNA Footprinting / methods*
  • Genomics
  • Micrococcal Nuclease / metabolism
  • Nucleosomes* / chemistry
  • Nucleosomes* / genetics
  • Nucleosomes* / metabolism
  • Saccharomyces cerevisiae / genetics
  • Sequence Analysis, DNA / methods*

Substances

  • Nucleosomes
  • DNA
  • Micrococcal Nuclease