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1.3K Views
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08:24 min
July 27th, 2021
DOI :
10.3791/61689-v
Chapters
0:04
Introduction
0:51
Cell Preparation
3:58
Proximity Ligation Assay (PLA)
6:20
Results: Representative Replisome Encounter Visualization
7:52
Conclusion
Transcript
该技术有助于可视化复制体抗原标记的DNA加合物遭遇,并且可以扩展到任何蛋白质与适合免疫检测的DNA结构或病变的相互作用。该方法可用于评估单个细胞内的相互作用频率,而不是整个群体,并且允许原位可视化细胞周期依赖性相互作用而不会损坏细胞。演示该程序的是Seidman实验室的博士后Ishani Majumdar。
在实验前一天,将1.5乘以10至2毫升培养基中的第五个细胞接种到用细胞粘合剂溶液预处理的35毫米玻璃底培养皿上。第二天,用37摄氏度5微摩尔Dig TMP补充培养基将每个培
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Summary
虽然与DNA加合物的复制叉碰撞可以诱导双链断裂,但对爬酶体和阻断病变之间的相互作用知之甚少。我们采用邻近连接测定来可视化这些遭遇并表征对重复体组成的影响。
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