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Optimization of experimental design parameters for high-throughput chromatin immunoprecipitation studies.

ABSTRACT: High-throughput, microarray-based chromatin immunoprecipitation (ChIP-chip) technology allows in vivo elucidation of transcriptional networks. However this complex is not yet readily accessible, in part because its many parameters have not been systematically evaluated and optimized. We address this gap by systematically assessing experimental-design parameters including antibody purity, dye-bias, array-batch, inter-day hybridization bias, amplification method and choice of hybridization control. The combined performance of these optimized parameters shows a 90% validation rate in ChIP-chip analysis of Myc genomic binding in HL60 cells using two different microarray platforms. Increased sensitivity and decreased noise in ChIP-chip assays will enable wider use of this methodology to accurately and affordably elucidate transcriptional networks.

SUBMITTER: Ponzielli R

PROVIDER: S-EPMC2588500 | biostudies-literature | 2008 Dec

REPOSITORIES: biostudies-literature

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Optimization of experimental design parameters for high-throughput chromatin immunoprecipitation studies.

Ponzielli Romina R Boutros Paul C PC Katz Sigal S Stojanova Angelina A Hanley Adam P AP Khosravi Fereshteh F Bros Christina C Jurisica Igor I Penn Linda Z LZ

Nucleic acids research 20081021 21

High-throughput, microarray-based chromatin immunoprecipitation (ChIP-chip) technology allows in vivo elucidation of transcriptional networks. However this complex is not yet readily accessible, in part because its many parameters have not been systematically evaluated and optimized. We address this gap by systematically assessing experimental-design parameters including antibody purity, dye-bias, array-batch, inter-day hybridization bias, amplification method and choice of hybridization control ...[more]

PMID: 18940864

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