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Biochemistry

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JoVE Journal Biochemistry
Monitoring Protein-RNA Interaction Dynamics In Vivo at High Temporal Resolution Using χCRAC
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Monitoring Protein-RNA Interaction Dynamics In Vivo at High Temporal Resolution Using χCRAC

Article DOI: 10.3791/61027-v • 09:15 min • May 9th, 2020
May 9th, 2020 •
1, 1, 2, 3, 1, 1, 4, 4, 3, 1
1Centre for Engineering Biology, University of Edinburgh, 2Institute of Cell Biology, University of Edinburgh, 3Department of Molecular Biology and Genetics, Aarhus University, 4UVO3 Ltd.

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Summary
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Kinetic cross-linking and analysis of cDNA is a method that allows investigation of the dynamics of protein-RNA interactions in living cells at high temporal resolution. Here the protocol is described in detail, including the growth of yeast cells, UV cross-linking, harvesting, protein purification, and next generation sequencing library preparation steps.

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Protein-RNA Interaction CRAC Temporal Resolution Live Cells Crosslinker UV Damage Responses Stress Responses RNA Degradation Visual Demonstration UV Crosslinking Harvesting Of Cells Extraction Of Crosslinked RNAs Microorganisms Solution Inoculation OD 600 Vari-X-linker Crosslinker Nanometer UV Vacuum Filtration Device
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