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【资料】An efficient ChIP protocol
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An efficient chromatin immunoprecipitation (ChIP) protocol for studying histone modifications in Arabidopsis plants 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 lg of DNA from each immunoprecipitated sample (from 0.2 to 0.4 g of starting tissue material) sufficient to test Z25 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.[ Last edited by silicare on 2012-11-1 at 10:44 ] |
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2012-10-08 15:44:54, 599.17 K
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5楼2012-11-01 10:28:52
7楼2013-07-13 10:12:19
8楼2014-08-07 14:13:58
简单回复
2012-10-08 16:45
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五星好评 感谢分享！！！
silicare3楼
2012-10-08 17:00
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五星好评 顶一下，感谢分享！
果壳4楼
2012-10-09 16:23
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五星好评 顶一下，感谢分享！
mlw09276楼
2013-01-05 11:36
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五星好评 顶一下，感谢分享！