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Related Experiment Videos

DNA Accessibility by MNase Digestions.

Ann-Kristin Östlund Farrants1

  • 1Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Svante Arrhenius väg 20C, 106 91, Stockholm, Sweden. anki.ostlund@su.se.

Methods in Molecular Biology (Clifton, N.J.)
|October 14, 2017
PubMed
Summary
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Micrococcal nuclease (MNase) digestion fragments chromatin to reveal nucleosome positions. Optimizing MNase titration is crucial for accurate genome-wide nucleosome mapping using sequencing or qPCR.

Area of Science:

  • Molecular Biology
  • Genomics
  • Epigenetics

Background:

  • Chromatin structure, organized by nucleosomes, plays a vital role in gene regulation.
  • Understanding nucleosome positioning is key to deciphering regulatory mechanisms.

Purpose of the Study:

  • To describe the method of using micrococcal nuclease (MNase) digestion for nucleosome mapping.
  • To highlight the importance of MNase titration for accurate chromatin analysis.

Main Methods:

  • Chromatin digestion using micrococcal nuclease (MNase) to generate mononucleosomes.
  • Analysis of protected DNA fragments via genome-wide sequencing or quantitative PCR (qPCR).

Main Results:

  • MNase digestion effectively isolates mononucleosomes by cleaving linker DNA.
Keywords:
Chromatin accessibilityMNase digestionNucleosome position

Related Experiment Videos

  • Differential sensitivity of nucleosomes to MNase requires careful titration for comprehensive mapping.

    • Conclusions:

    • MNase digestion is a powerful technique for mapping nucleosome positions across the genome.
    • Optimized MNase titration is essential for generating high-resolution chromatin maps.