STAR Protocols (Jun 2021)

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

  • Laura E. McKnight,
  • Johnathan G. Crandall,
  • Thomas B. Bailey,
  • Orion G.B. Banks,
  • Kona N. Orlandi,
  • Vi N. Truong,
  • Drake A. Donovan,
  • Grace L. Waddell,
  • Elizabeth T. Wiles,
  • Scott D. Hansen,
  • Eric U. Selker,
  • Jeffrey N. McKnight

Journal volume & issue
Vol. 2, no. 2
p. 100486

Abstract

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Summary: 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.

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