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使用色素免疫沉淀-外核酶（ChIP-Exo）对小鼠干细胞衍生神经元中的蛋白质-DNA相互作用进行高分辨率映射

JoVE Journal
Genética

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JoVE Journal Genética

High-Resolution Mapping of Protein-DNA Interactions in Mouse Stem Cell-Derived Neurons using Chromatin Immunoprecipitation-Exonuclease (ChIP-Exo)

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使用色素免疫沉淀-外核酶（ChIP-Exo）对小鼠干细胞衍生神经元中的蛋白质-DNA相互作用进行高分辨率映射

DOI:

10.3791/61124-v

•

08:40 min

•

August 14, 2020

•

Kaitlin N. Montanera², Ho Sung Rhee²

¹Department of Cell and Systems Biology,University of Toronto, ²Department of Biology,University of Toronto

Capítulos

00:05Introduction
00:21Antibody Incubation with Beads
01:35Chromatin Immunoprecipitation (ChIP)
02:56Enzymatic Reactions on Beads
04:59Elution and Purification
05:43Library Preparation
06:50Results: ChIP-exo Produces High Mapping Resolution
08:04Conclusion

Summary

Tadução automática

精确确定整个基因组中的蛋白质结合位置对于理解基因调控非常重要。在这里，我们描述了一种基因组映射方法，它用外核酶消化（ChIP-exo）处理染色质免疫沉淀DNA，然后进行高通量测序。该方法可检测哺乳动物神经元中具有近碱基对映射分辨率和高信号噪声比的蛋白质-DNA相互作用。

Tags

Keywords: ChIP-exo Protein-DNA Interactions Chromatin Immunoprecipitation High-resolution Mapping Mouse Stem Cell-derived Neurons Magnetic Beads Blocking Solution Islet-1 Antibody Sonicated Lysates Wash Buffer End Prep Reaction Mix End Prep Enzyme Mix

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