高分辨率染色质免疫沉淀：ChIP测序

High-Resolution Chromatin Immunoprecipitation: ChIP-Sequencing.

作者信息

Diaz Roxanne E, Sanchez Aurore, Anton Le Berre Véronique, Bouet Jean-Yves

机构信息

Laboratoire de Microbiologie et Génétique Moléculaires, Centre de Biologie Intégrative (CBI), Centre National de la Recherche Scientifique (CNRS), Université de Toulouse, UPS, F-31000, Toulouse, France.

Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, Université de Toulouse, UPS, INSA, INP, CNRS, F-31077, Toulouse, France.

出版信息

Methods Mol Biol. 2017;1624:61-73. doi: 10.1007/978-1-4939-7098-8_6.

DOI:10.1007/978-1-4939-7098-8_6

PMID:28842876

Abstract

Chromatin immunoprecipitation (ChIP) coupled with next-generation sequencing (NGS) is widely used for studying the nucleoprotein components that are involved in the various cellular processes required for shaping the bacterial nucleoid. This methodology, termed ChIP-sequencing (ChIP-seq), enables the identification of the DNA targets of DNA binding proteins across genome-wide maps. Here, we describe the steps necessary to obtain short, specific, high-quality immunoprecipitated DNA prior to DNA library construction for NGS and high-resolution ChIP-seq data.

摘要

染色质免疫沉淀（ChIP）与下一代测序（NGS）相结合，被广泛用于研究参与塑造细菌类核所需的各种细胞过程的核蛋白成分。这种方法称为ChIP测序（ChIP-seq），能够在全基因组图谱上识别DNA结合蛋白的DNA靶点。在这里，我们描述了在构建用于NGS的DNA文库和获得高分辨率ChIP-seq数据之前，获得短的、特异性的、高质量免疫沉淀DNA所需的步骤。