Microplate-based chromatin immunoprecipitation method, Matrix ChIP: a platform to study signaling of complex genomic events

被引:58
作者
Flanagin, Steve [1 ]
Nelson, Joel D. [2 ]
Castner, David G. [3 ]
Denisenko, Oleg [1 ]
Bomsztyk, Karol [1 ]
机构
[1] Univ Washington, Dept Med, UW Med Lake Union, Seattle, WA 98109 USA
[2] Univ Washington, Mol & Cellular Biol Program, Seattle, WA 98109 USA
[3] Univ Washington, Dept Bioengn & Chem Engn, Natl ESCA & Surface Anal Ctr, Biomed Program, Seattle, WA 98195 USA
关键词
D O I
10.1093/nar/gkn001
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 [生物化学与分子生物学]; 081704 [应用化学];
摘要
The chromatin immunoprecipitation (ChIP) assay is a major tool in the study of genomic processes in vivo. This and other methods are revealing that control of gene expression, cell division and DNA repair involves multiple proteins and great number of their modifications. ChIP assay is traditionally done in test tubes limiting the ability to study signaling of the complex genomic events. To increase the throughput and to simplify the assay we have developed a microplate-based ChIP (Matrix ChIP) method, where all steps from immunoprecipitation to DNA purification are done in microplate wells without sample transfers. This platform has several important advantages over the tube-based assay including very simple sample handling, high throughput, improved sensitivity and reproducibility, and potential for automation. 96 ChIP measurements including PCR can be done by one researcher in one day. We illustrate the power of Matrix ChIP by parallel profiling 80 different chromatin and transcription time-course events along an inducible gene including transient recruitment of kinases.
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页数:9
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