Reimer Julia J, Turck Franziska
Max Planck Institute for Plant Breeding Research, Köln, Germany.
Methods Mol Biol. 2010;631:139-60. doi: 10.1007/978-1-60761-646-7_12.
Chromatin immunoprecipitation in combination with DNA-microarray hybridization (ChIP-chip) allows the identification of chromatin regions that are associated with modified forms of histones on a genomic scale. The ChIP-chip workflow consists of the following steps: generation of biological material, in vivo formaldehyde-fixation of protein-DNA and protein-protein interactions, chromatin preparation and shearing, immunoprecipitation of chromatin with specific antibodies, fixation reversal and DNA purification, DNA amplification, microarray hybridization, and data analysis. In Part A of this chapter, we describe molecular methods of the experimental procedure employed to identify chromosomal regions of Arabidopsis thaliana associated with H3K27me3. In addition, some general information on the microarray platform from Roche-NimbleGen will be provided. Part B of this chapter focuses on ChIP-chip data analysis of H3K27me3 on the Roche-NimbleGen platform.
染色质免疫沉淀结合DNA微阵列杂交（ChIP-chip）技术能够在全基因组范围内鉴定与组蛋白修饰形式相关的染色质区域。ChIP-chip工作流程包括以下步骤：生物材料的制备、蛋白质-DNA和蛋白质-蛋白质相互作用的体内甲醛固定、染色质制备与剪切、用特异性抗体进行染色质免疫沉淀、固定逆转与DNA纯化、DNA扩增、微阵列杂交以及数据分析。在本章的A部分，我们将描述用于鉴定拟南芥中与H3K27me3相关的染色体区域的实验过程的分子方法。此外，还将提供有关罗氏- NimbleGen微阵列平台的一些一般信息。本章的B部分重点介绍罗氏- NimbleGen平台上H3K27me3的ChIP-chip数据分析。
