An Optimized Chromatin Immunoprecipitation Protocol for Quantification of Protein-DNA Interactions

一种用于定量分析蛋白质-DNA相互作用的优化染色质免疫沉淀方案

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作者:Wim J de Jonge ,Mariël Brok ,Patrick Kemmeren ,Frank C P Holstege

Abstract

Transcription factors are important regulators of cell fate and function. Knowledge about where transcription factors are bound in the genome is crucial for understanding their function. A common method to study protein-DNA interactions is chromatin immunoprecipitation (ChIP). Here, we present a revised ChIP protocol to determine protein-DNA interactions for the yeast Saccharomyces cerevisiae. We optimized several aspects of the procedure, including cross-linking and quenching, cell lysis, and immunoprecipitation steps. This protocol facilitates sensitive and reproducible quantitation of protein-DNA interactions. For complete details on the use and execution of this protocol, please refer to (de Jonge et al., 2019).

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