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

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

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4,587 Views

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09:15 min

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May 09, 2020

DOI:

10.3791/61027-v

DOI:
10.3791/61027-v

•

09:15 min
May 09, 2020

•

33 Views
, , , , , , , , ,
1Centre for Engineering Biology,University of Edinburgh, 2Institute of Cell Biology,University of Edinburgh, 3Department of Molecular Biology and Genetics,Aarhus University, 4UVO3 Ltd.

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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Cite this Article

McKellar, S. W., Ivanova, I., van Nues, R. W., Cordiner, R. A., Chauhan, M., Christopoulou, N., Worboys, W., Langford, A., Jensen, T. H., Granneman, S. Monitoring Protein-RNA Interaction Dynamics In Vivo at High Temporal Resolution Using χCRAC. J. Vis. Exp. (159), e61027, doi:10.3791/61027 (2020).

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