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快速染色质免疫沉淀测定法。

Fast chromatin immunoprecipitation assay.

作者信息

Nelson Joel D, Denisenko Oleg, Sova Pavel, Bomsztyk Karol

机构信息

Molecular and Cellular Biology Program, University of Washington, Seattle, WA 98109, USA.

出版信息

Nucleic Acids Res. 2006 Jan 5;34(1):e2. doi: 10.1093/nar/gnj004.

Abstract

Chromatin immunoprecipitation (ChIP) is a widely used method to explore in vivo interactions between proteins and DNA. The ChIP assay takes several days to complete, involves several tube transfers and uses either phenol-chlorophorm or spin columns to purify DNA. The traditional ChIP method becomes a challenge when handling multiple samples. We have developed an efficient and rapid Chelex resin-based ChIP procedure that dramatically reduces time of the assay and uses only a single tube to isolate PCR-ready DNA. This method greatly facilitates the probing of chromatin changes over many time points with several antibodies in one experiment.

摘要

染色质免疫沉淀(ChIP)是一种广泛用于探索蛋白质与DNA在体内相互作用的方法。ChIP分析需要几天时间才能完成,涉及多次试管转移,并使用酚-氯仿或离心柱来纯化DNA。在处理多个样本时,传统的ChIP方法成为一项挑战。我们开发了一种基于Chelex树脂的高效快速ChIP程序,该程序显著缩短了分析时间,并且仅使用单个试管来分离可用于PCR的DNA。这种方法极大地便于在一个实验中使用几种抗体在多个时间点探测染色质变化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97bb/1325209/dd44d168487f/gnj004f1.jpg

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