Hi-C 2.0: An optimized Hi-C procedure for high-resolution genome-wide mapping of chromosome conformation.

Hi-C 2.0: An optimized Hi-C procedure for high-resolution genome-wide mapping of chromosome conformation.
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DOI:
10.1016/j.ymeth.2017.04.004
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发表时间:
2017-07-01
期刊:
Methods (San Diego, Calif.)
影响因子:
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通讯作者:
Gibcus JH
Gibcus JH
中科院分区:
其他
文献类型:
--
作者:
Belaghzal H;Dekker J;Gibcus JH

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基于染色体构象捕获的方法如Hi-C已经成为研究基因组三维结构的主流技术。这些方法将反映拓扑染色质结构的染色质相互作用转换为数字信息(成对相互作用的计数)。在这里,我们描述了一个更新的协议Hi-C(Hi-C 2.0),它集成了最近的改进到一个单一的协议,高效和高分辨率的染色质相互作用的捕获。该方案将染色质消化和频繁切割酶结合以获得酶解(Kb)。它还包括减少随机连接和无信息分子(例如未连接末端)的产生的步骤,以提高有效染色体内读段对的量。该方案允许获得关于构象结构的信息,例如区室和拓扑相关结构域,以及高分辨率构象特征,例如DNA环。
Chromosome conformation capture-based methods such as Hi-C have become mainstream techniques for the study of the 3D organization of genomes. These methods convert chromatin interactions reflecting topological chromatin structures into digital information (counts of pair-wise interactions). Here, we describe an updated protocol for Hi-C (Hi-C 2.0) that integrates recent improvements into a single protocol for efficient and high-resolution capture of chromatin interactions. This protocol combines chromatin digestion and frequently cutting enzymes to obtain kilobase (Kb) resolution. It also includes steps to reduce random ligation and the generation of uninformative molecules, such as unligated ends, to improve the amount of valid intra-chromosomal read pairs. This protocol allows for obtaining information on conformational structures such as compartment and topologically associating domains, as well as high-resolution conformational features such as DNA loops.
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