Profiling Nucleosome Occupancy by MNase-seq: Experimental Protocol and Computational Analysis

Methods Mol Biol. 2018:1675:167-181. doi: 10.1007/978-1-4939-7318-7_11.

Abstract

Nucleosomes are the basic repeating units of eukaryotic chromatin. They play important roles in chromatin compaction and gene regulation. Therefore, it is important to profile the in vivo locations of nucleosomes in the genome. Here we illustrate how to profile nucleosome occupancy at genome-wide scale using micrococcal nuclease (MNase) digestion combined with high throughput Illumina sequencing (MNase-seq). Nucleosome-associated DNA is relatively insensitive to digestion by micrococcal nuclease (MNase). Upon mild MNase treatment, the undigested nucleosomal DNA can be purified and sequenced allowing a precise localization of in vivo nucleosomes at a genome-wide level.

Keywords: Chromatin; MNase digestion; MNase-seq; Next-generation sequencing; Nucleosome occupancy.

MeSH terms

  • Arabidopsis / genetics*
  • Chromosome Mapping
  • Computational Biology / methods
  • DNA, Plant
  • Gene Expression Regulation, Plant
  • High-Throughput Nucleotide Sequencing / methods*
  • Micrococcal Nuclease / metabolism
  • Nucleosomes / genetics*
  • Sequence Analysis, DNA / methods*

Substances

  • DNA, Plant
  • Nucleosomes
  • Micrococcal Nuclease