Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai 200032, China.
Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine and Science, MN 55905, USA.
STAR Protoc. 2021 Jul 17;2(3):100681. doi: 10.1016/j.xpro.2021.100681. eCollection 2021 Sep 17.
Inhibition of histone deacetylases causes rapid and robust acetylation of histones. In this case, histone acetylation is likely increased on nearly every nucleosome, and the per-cell DNA/chromatin yield in chromatin immunoprecipitation (ChIP) experiments is significantly increased. Spike-in controls are essential for normalizing ChIP sequencing (ChIP-seq) data to capture this massive effect. Here, we report a detailed protocol of H3K27-ac ChIP-seq in human cells with chromatin from an ancestral species as a spike-in control. For complete details on the use and execution of this protocol, please refer to Wu et al. (2021).
组蛋白去乙酰化酶的抑制作用会导致组蛋白迅速而强烈的乙酰化。在这种情况下，组蛋白乙酰化可能几乎在每个核小体上都增加，并且染色质免疫沉淀（ChIP）实验中的每个细胞的 DNA/染色质产量显著增加。 Spike-in 对照对于将 ChIP 测序（ChIP-seq）数据标准化以捕获这种巨大影响是必不可少的。在这里，我们报告了一种使用祖先物种的染色质作为 Spike-in 对照进行人细胞中 H3K27ac ChIP-seq 的详细方案。有关此方案的使用和执行的完整详细信息，请参阅 Wu 等人。（2021 年）。
