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Journal
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Biochemistry
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使用χCRAC以高时间分辨率监测体内蛋白质-RNA相互作用动力学
JoVE Journal
Biochemistry
This content is Free Access.
JoVE Journal
Biochemistry
Monitoring Protein-RNA Interaction Dynamics In Vivo at High Temporal Resolution Using χCRAC
Please note that all translations are automatically generated.
Click here for the English version.
使用χCRAC以高时间分辨率监测体内蛋白质-RNA相互作用动力学
DOI:
10.3791/61027-v
•
09:15 min
•
May 09, 2020
•
Stuart W. McKellar
,
Ivayla Ivanova
,
Robert W. van Nues
,
Ross A. Cordiner
,
Mehak Chauhan
,
Niki Christopoulou
,
Will Worboys
,
Andrew Langford
,
Torben Heick Jensen
,
Sander Granneman
1
Centre for Engineering Biology
,
University of Edinburgh
,
2
Institute of Cell Biology
,
University of Edinburgh
,
3
Department of Molecular Biology and Genetics
,
Aarhus University
,
4
UVO3 Ltd.
Chapters
00:04
Introduction
00:58
UV Crosslinking and Lysate Production
03:18
Elution and SDS-PAGE
05:24
PCR and Gel Extraction
07:06
Results: Temporal Binding of Nrd1 to its Targets
08:47
Conclusion
Summary
Automatic Translation
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Automatic Translation
cDNA的动力学交联和分析是一种允许以高时间分辨率研究活细胞中蛋白质-RNA相互作用动力学的方法。这里详细描述了该方案,包括酵母细胞的生长、UV交联、收获、蛋白质纯化和下一代测序文库的制备步骤。
Tags
Protein-RNA Interaction
Kinetic CRAC
UV Crosslinking
RNA Binding Proteins
RNA Extraction
Cell Harvesting
Time-resolved Analysis
Live Cell Monitoring
Stress Response
Adherent Cells
Microorganisms
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