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Analyse van de beëindiging van de transcriptie gebruik van BrUTP-streng-specifieke transcriptie Run-on (TRO) Approach
JoVE Journal
Genetics
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JoVE Journal Genetics
Analysis of Termination of Transcription Using BrUTP-strand-specific Transcription Run-on (TRO) Approach
DOI:

12:12 min

March 12, 2017

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Chapters

  • 00:05Title
  • 01:39Transcription Run-on Reaction
  • 02:36RNA Extraction
  • 05:28Affinity Purification of BrUTP-labeled RNA
  • 07:56Precipitation of BrUTP-labeled RNA
  • 09:23Results: Rna14 is Required for Termination of Transcription of ASC1
  • 11:00Conclusion

Summary

Automatic Translation

We describe a basic experimental approach for analysis of termination of transcription by RNA polymerase II in vivo using BrUTP by the strand-specific transcription run-on (TRO) approach in budding yeast. This protocol can be extended to study transcription termination by other RNA polymerases both in yeast and higher eukaryotes.

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