Abstract
MNase-Seq is a genome-wide procedure that allows mapping of DNA associated to nucleosomes following micrococcal nuclease digestion. It is a rapid and robust technology useful for the analysis of chromatin properties genome-wide at the resolution of mono-nucleosomes. Here, we describe how to produce high-resolution nucleosome maps of cells grown in suspension or adherent mammalian cells. After only three steps: nuclei or cell preparation, native MNase digestion and DNA purification, libraries for high-throughput sequencing can be prepared. Genome-wide nucleosome maps allow analyzing chromatin opening at promoters or enhancers, nucleosome displacement, or labile nucleosome occupancy depending on the digestion condition used. As presented, MNase-Seq is a versatile tool for investigating chromatin dynamics, regulation, and to define open chromatin regions of regulatory elements in mammalian genomes.
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Acknowledgments
This work was supported by the French National Research Agency (ANR, ANR-18-CE12-0020) and the CNRS-UMR 5535 Institut de Génétique Moléculaire de Montpellier.
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Esnault, C., Magat, T., García-Oliver, E., Andrau, JC. (2021). Analyses of Promoter , Enhancer, and Nucleosome Organization in Mammalian Cells by MNase-Seq. In: Borggrefe, T., Giaimo, B.D. (eds) Enhancers and Promoters. Methods in Molecular Biology, vol 2351. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1597-3_5
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DOI: https://doi.org/10.1007/978-1-0716-1597-3_5
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