Princess Máxima Center for Pediatric Oncology, Heidelberglaan 25, 3584 CS Utrecht, the Netherlands.
STAR Protoc. 2020 Jun 19;1(1):100020. doi: 10.1016/j.xpro.2020.100020.
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 . 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).
转录因子是细胞命运和功能的重要调节剂。了解转录因子在基因组中的结合位置对于理解其功能至关重要。研究蛋白质-DNA 相互作用的一种常用方法是染色质免疫沉淀 (ChIP)。在这里,我们提出了一个修改后的酵母 ChIP 方案,以确定蛋白质-DNA 相互作用。我们优化了该程序的几个方面,包括交联和淬灭、细胞裂解和免疫沉淀步骤。该方案促进了蛋白质-DNA 相互作用的敏感和可重复定量。有关此方案的使用和执行的完整详细信息,请参考 (de Jonge 等人,2019 年)。