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MNase-Seq Analysis for Identifying Stress-Altered Nucleosome Occupancy in Plants

  • Caleb Watkins,
  • Karyn L. Willyerd,
  • Chi-ping Liao,
  • Donald R. Ruhl,
  • Charles Chen

摘要

Nucleosome occupancy plays an important role in chromatin compaction, affecting biological processes by hampering the binding of cis-acting elements such as transcription factors, RNA polymerase machinery, and coregulatory. Accessible regions allow for cis-acting elements to bind DNA and regulate transcription. Here, we detail our protocol to profile nucleosome occupancy and chromatin structure dynamics under drought stress at the genome-wide scale using micrococcal nuclease (MNase) digestion. Combining variable MNase concentration treatments and high-throughput sequencing, we investigate the changes in the overall chromatin state using bread wheat samples from an exemplary drought experiment.