Protocol for the fast chromatin immunoprecipitation (ChIP) method.
Chromatin and transcriptional processes are among the most intensively studied fields of biology today. The introduction of chromatin immunoprecipitations (ChIP) represents a major advancement in this area. This powerful method allows researchers to probe specific protein-DNA interactions in vivo and to estimate the density of proteins at specific sites genome-wide. We have introduced several improvements to the traditional ChIP assay, which simplify the procedure, greatly reducing the time and labor required to complete the assay. The simplicity of the method yields highly reproducible results. Our improvements facilitate the probing of multiple proteins in a single experiment, which allows for the simultaneous monitoring of many genomic events. This method is particularly useful in kinetic studies where multiple samples are processed at the same time. Starting with sheared chromatin, PCR-ready DNA can be isolated from 16-24 ChIP samples in 4-6 h using the fast method.
Nelson
Joel D
JD
Molecular and Cellular Biology Program, University of Washington, Seattle, Washington 98109, USA.
Denisenko
Oleg
O
Bomsztyk
Karol
K
D002843:Chromatin / Q000737:chemistry; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D004268:DNA-Binding Proteins / Q000378:metabolism
D002843:Chromatin; D004268:DNA-Binding Proteins