Department of Medicine, Division of Hematology-Oncology and Cancer Research Institute, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02215, USA.
Department of Medicine, Division of Hematology-Oncology and Cancer Research Institute, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02215, USA.
STAR Protoc. 2022 Jul 31;3(3):101529. doi: 10.1016/j.xpro.2022.101529. eCollection 2022 Sep 16.
Chromatin immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) even with optimization may give low signal-to-background ratio and spatial resolution. Here, we adapted Cleavage Under Targets and Release Using Nuclease (CUT&RUN) (originally developed by the Henikoff group) to develop CUT&RUN-qPCR. By studying the recruitment of selected proteins (but amenable to other proteins), we find that CUT&RUN-qPCR is more sensitive and gives better spatial resolution than ChIP-qPCR. For complete details on the use and execution of this protocol, please refer to Skene et al. (2018) and Skene and Henikoff (2017).
染色质免疫沉淀结合定量 PCR（ChIP-qPCR）即使经过优化，也可能产生低信号背景比和空间分辨率。在这里，我们改编了靶向切割和释放使用核酸酶（CUT&RUN）（最初由 Henikoff 小组开发）来开发 CUT&RUN-qPCR。通过研究选定蛋白质的募集（但适用于其他蛋白质），我们发现 CUT&RUN-qPCR 比 ChIP-qPCR 更灵敏，空间分辨率更好。有关此方案的使用和执行的完整详细信息，请参阅 Skene 等人。（2018 年）和 Skene 和 Henikoff（2017 年）。
