胚胎
劈理(地质)
卵母细胞
细胞生物学
免疫沉淀
刀豆蛋白A
核酸酶
染色质
DNA
化学
生物
分子生物学
抗体
遗传学
生物化学
体外
古生物学
断裂(地质)
作者
Ryoya Hayashi,Azusa Inoue
标识
DOI:10.1007/978-1-0716-2724-2_6
摘要
Cleavage Under Target & Release Using Nuclease (CUT&RUN) enables the detection of DNA regions that are bound by a protein of interest. This method is suitable for low-input materials because of the absence of an immunoprecipitation step. However, it sometimes fails when applying it to fragile cells, such as mouse oocytes. Here we describe our low-input CUT&RUN protocol optimized for mouse oocyte and preimplantation embryo samples in which the primary antibody and protein A-MNase binding steps are completed before the cells are bound to Concanavalin A-coated magnetic beads. This modification prevents crush of oocytes and early embryos and unwanted loss of chromatin during CUT&RUN procedures.
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