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Nat Protoc. 2008;3(6):1018-25. doi: 10.1038/nprot.2008.66.

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

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  • 1School of Biological Sciences, University of Nebraska, Lincoln, Nebraska 68588, USA. asaleh@ncsu.edu

Abstract

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 mug of DNA from each immunoprecipitated sample (from 0.2 to 0.4 g of starting tissue material) sufficient to test > or =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.

PMID:
18536649
[PubMed - indexed for MEDLINE]
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