Quantitative Cell Biology Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
STAR Protoc. 2020 Sep 18;1(2):100062. doi: 10.1016/j.xpro.2020.100062.
Chromatin immunoprecipitation (ChIP) followed by next-generation sequencing is a powerful technique that characterizes the genome-wide DNA-binding profile of a protein of interest. The general ChIP-seq workflow has been applied widely to many sample types and target proteins, but sample-specific optimization of various steps is necessary to achieve high-quality data. This protocol is specifically optimized for cultured human embryonic stem cells (hESCs), including steps to check sample quality and non-specific enrichment of "hyper-ChIPable" regions prior to sequencing. For complete details on the use and execution of this protocol, please refer to Gunne-Braden et al. (2020).
染色质免疫沉淀(ChIP)结合下一代测序是一种强大的技术,可描绘目标蛋白质的全基因组 DNA 结合图谱。一般的 ChIP-seq 工作流程已广泛应用于许多样本类型和靶蛋白,但为了获得高质量的数据,需要对各个步骤进行样本特异性优化。本方案专门针对培养的人胚胎干细胞(hESC)进行了优化,包括在测序前检查样品质量和“高 ChIP 可及性”区域非特异性富集的步骤。有关本方案的使用和执行的完整详细信息,请参阅 Gunne-Braden 等人(2020 年)。