使用直接核小体条形码的多重染色质免疫沉淀测序：一种用于高通量染色质分析的工具

Multiplexed ChIP-Seq Using Direct Nucleosome Barcoding: A Tool for High-Throughput Chromatin Analysis.

作者信息

Chabbert Christophe D, Adjalley Sophie H, Steinmetz Lars M, Pelechano Vicent

机构信息

Genome Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.

Institute of Molecular Health Sciences, ETH Zurich, Zürich, 8093, Switzerland.

出版信息

Methods Mol Biol. 2018;1689:177-194. doi: 10.1007/978-1-4939-7380-4_16.

DOI:10.1007/978-1-4939-7380-4_16

PMID:29027175

Abstract

Chromatin immunoprecipitation followed by sequencing (ChIP-Seq) or microarray hybridization (ChIP-on-chip) are standard methods for the study of transcription factor binding sites and histone chemical modifications. However, these approaches only allow profiling of a single factor or protein modification at a time.In this chapter, we present Bar-ChIP, a higher throughput version of ChIP-Seq that relies on the direct ligation of molecular barcodes to chromatin fragments. Bar-ChIP enables the concurrent profiling of multiple DNA-protein interactions and is therefore amenable to experimental scale-up, without the need for any robotic instrumentation.

摘要

染色质免疫沉淀测序（ChIP-Seq）或微阵列杂交（芯片上的ChIP）是研究转录因子结合位点和组蛋白化学修饰的标准方法。然而，这些方法一次只能对单个因子或蛋白质修饰进行分析。在本章中，我们介绍了Bar-ChIP，这是一种ChIP-Seq的高通量版本，它依赖于将分子条形码直接连接到染色质片段上。Bar-ChIP能够同时对多种DNA-蛋白质相互作用进行分析，因此适合扩大实验规模，而无需任何机器人仪器。