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Analysis of RNA Processing Reactions Using Cell Free Systems: 3′ End Cleavage of Pre-mRNA Substrates in vitro
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Analysis of RNA Processing Reactions Using Cell Free Systems: 3′ End Cleavage of Pre-mRNA Substrates in vitro

Analysis of RNA Processing Reactions Using Cell Free Systems: 3′ End Cleavage of Pre-mRNA Substrates in vitro

DOI:
10.3791/51309-v

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09:16 min

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May 03, 2014

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, , ,
1Department of Infectious Diseases,The Scripps Research Institute, 2Department of Chemistry,City College of New York

Chapters

  • 00:05Title
  • 01:11Collecting Nuclear Extract
  • 04:09Probe Preparation and PAGE Extraction
  • 05:293′ mRNA Cleavage Reaction and PAGE Analysis
  • 08:04Cleavage Assay of RNA poly(A) for HIV-1
  • 08:44Conclusion

Summary

Automatic Translation

Automatic Translation

RNA polymerase II synthesizes a precursor RNA that extends beyond the 3' end of the mature mRNA. The end of the mature RNA is generated cotranscriptionally, at a site dictated by RNA sequences, via the endonuclease activity of the cleavage complex. Here, we detail the method to study cleavage reactions in vitro.

Tags

Keywords: RNA Processing3′ End CleavagePre-mRNACell-free SystemsIn VitroRNA Polymerase IIEndonuclease ActivityCleavage ComplexPolyadenylationNuclear ExtractsRadiolabeled RNA SubstratesHIV-1 MRNAs

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