An efficient chromatin immunoprecipitation (ChIP) protocol for studying histone modifications in Arabidopsis plants

被引：504

作者：

Saleh, Abdelaty ^{[1
,2
]}

Alvarez-Venegas, Raul ^{[1
,3
]}

Avramova, Zoya ^{[1
]}

机构：

[1] Univ Nebraska, Sch Biol Sci, Lincoln, NE 68588 USA

[2] N Carolina State Univ, Dept Plant Pathol, CIFR, Raleigh, NC 27695 USA

[3] Ctr Invest & Estud Avanzados, Dept Genet Engn, Irapuato 36821, Mexico

来源：

NATURE PROTOCOLS | 2008年 / 3卷 / 06期

关键词：

D O I：

10.1038/nprot.2008.66

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Chromatin immunoprecipitation (ChIP) is a powerful tool for the characterization of covalent histone modifications and DNA-histone interactions in vivo. The procedure includes DNA-histone cross-linking in chromatin, shearing DNA into smaller fragments, immunoprecipitation with antibodies against the histone modifications of interest, followed by PCR identification of associated DNA sequences. In this protocol, we describe a simplified and optimized version of ChIP assay by reducing the number of experimental steps and isolation solutions and shortening preparation times. We include a nuclear isolation step before chromatin shearing, which provides a good yield of high-quality DNA resulting in at least 15 mu g of DNA from each immunoprecipitated sample (from 0.2 to 0.4 g of starting tissue material) sufficient to test >= 25 genes of interest. This simpler and cost-efficient protocol has been applied for histone-modification studies of various Arabidopsis thaliana tissues and is easy to adapt for other systems as well.

引用

页码：1018 / 1025

页数：8

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