Cui Kairong, Zhao Keji
Laboratory of Molecular Immunology, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD, USA.
Methods Mol Biol. 2012;833:413-9. doi: 10.1007/978-1-61779-477-3_24.
The precise location of nucleosomes in functional regulatory regions in chromatin is critical to the regulation of transcription. The nucleosome structure protects DNA from microccocal nuclease (MNase) digestion and leaves a footprint on DNA that indicates the position of nucleosomes. Short sequence reads (25-36 bp) from ends of mononucleosome-sized DNA generated from MNase digestion of chromatin can be determined using next-generation sequencing techniques. Mapping of these short reads to the genome provides a powerful genome-wide approach to precisely define the nucleosome positions in any genome with known genomic sequence. This chapter outlines the reagents and experimental procedures of MNase-Seq for mapping nucleosome positions in the human genome.
染色质功能调控区域中核小体的精确位置对于转录调控至关重要。核小体结构可保护DNA免受微球菌核酸酶（MNase）消化，并在DNA上留下表明核小体位置的足迹。使用下一代测序技术可以确定由染色质MNase消化产生的单核小体大小DNA末端的短序列读数（25 - 36 bp）。将这些短读数定位到基因组提供了一种强大的全基因组方法，可精确确定任何具有已知基因组序列的基因组中的核小体位置。本章概述了用于绘制人类基因组中核小体位置的MNase-Seq试剂和实验程序。
