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通过微球菌核酸酶测序分析核小体占据情况:实验方案与计算分析

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

作者信息

Pajoro Alice, Muiño Jose M, Angenent Gerco C, Kaufmann Kerstin

机构信息

Laboratory of Molecular Biology, Wageningen University and Research, Droevendaalsesteeg 1, 6708 PB, Wageningen, The Netherlands.

Department of Computational Molecular Biology, Max-Planck Institute for Molecular Genetics, Ihnestraße 63-73, 14195, Berlin, Germany.

出版信息

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.

摘要

核小体是真核染色质的基本重复单元。它们在染色质压缩和基因调控中发挥重要作用。因此,描绘基因组中核小体的体内位置非常重要。在这里,我们阐述了如何使用微球菌核酸酶(MNase)消化结合高通量Illumina测序(MNase-seq)在全基因组范围内描绘核小体占有率。与核小体相关的DNA对微球菌核酸酶(MNase)的消化相对不敏感。经过温和的MNase处理后,未消化的核小体DNA可以被纯化并测序,从而在全基因组水平上精确定位体内核小体。

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