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Monitoring Protein-RNA Interaction Dynamics In Vivo at High Temporal Resolution Using χCRAC
JoVE Journal
Biochemistry
This content is Free Access.
JoVE Journal Biochemistry
Monitoring Protein-RNA Interaction Dynamics In Vivo at High Temporal Resolution Using χCRAC
DOI:

09:15 min

May 09, 2020

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Chapters

  • 00:04Introduction
  • 00:58UV Crosslinking and Lysate Production
  • 03:18Elution and SDS-PAGE
  • 05:24PCR and Gel Extraction
  • 07:06Results: Temporal Binding of Nrd1 to its Targets
  • 08:47Conclusion

Summary

Automatic Translation

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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