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生物化学
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Extremely Rapid and Specific Metabolic Labelling of RNA In Vivo with 4-Thiouracil (Ers4tU)
JoVE 杂志
生物化学
This content is Free Access.
JoVE 杂志 生物化学
Extremely Rapid and Specific Metabolic Labelling of RNA In Vivo with 4-Thiouracil (Ers4tU)

Extremely Rapid and Specific Metabolic Labelling of RNA In Vivo with 4-Thiouracil (Ers4tU)

DOI:
10.3791/59952-v

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00:11 min

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August 22, 2019

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,
1Wellcome Centre for Cell Biology, School of Biological Sciences,University of Edinburgh

Chapters

  • 00:04Title
  • 00:38Growth and Thio-labeling
  • 02:55Preparation of Total RNA
  • 04:38Biotinylation
  • 05:58Purification of the Newly Synthesized RNA
  • 09:29Results: In Vivo Labelling of RNA with 4-Thiouracil
  • 10:44Conclusion

Summary

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The use of thiolated uracil to sensitively and specifically purify newly transcribed RNA from the yeast Saccharomyces cerevisiae.

Tags

Keywords: 4-ThiouracilRNA LabelingIn VivoPulse-chase ExperimentsProtein DepletionThiolationYeastS. PombeMetabolic LabelingRNA ExtractionAE BufferPhenol Extraction

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