Department of Medical Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan.
Research Laboratory, H.U. Group Research Institute G.K., Tokyo, Japan.
Methods Mol Biol. 2023;2577:55-64. doi: 10.1007/978-1-0716-2724-2_4.
Chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) is the most widely used method for analyzing genome-wide DNA-protein interactions. Because there is considerable variation in the modes and strengths of DNA-protein interactions, chromatin immunoprecipitation (ChIP) protocols have been diversified and optimized for different needs. Here, we describe protocols for detecting histone modifications and modifiers using various crosslinking and immunoprecipitation conditions. We provide a complete ChIP-seq workflow covering sample preparation, immunoprecipitation, next-generation sequencing (NGS) library preparation, and data analyses.
染色质免疫沉淀结合高通量测序(ChIP-seq)是分析全基因组 DNA-蛋白质相互作用最广泛使用的方法。由于 DNA-蛋白质相互作用的模式和强度存在很大的差异,因此已经针对不同的需求对染色质免疫沉淀(ChIP)方案进行了多样化和优化。在这里,我们描述了使用各种交联和免疫沉淀条件检测组蛋白修饰和修饰剂的方案。我们提供了一个完整的 ChIP-seq 工作流程,涵盖了样品制备、免疫沉淀、下一代测序(NGS)文库制备和数据分析。