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Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution
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Biochemistry
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JoVE Journal Biochemistry
Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution

Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution

DOI:
10.3791/54391-v

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10:53 min

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January 16, 2017

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, , , ,
1Department of Veterinary & Biomedical Sciences,University of Minnesota, 2Department of Veterinary Biosciences,Ohio State University, 3School of Biotechnology,Gautam Buddha University

Chapters

  • 00:05Title
  • 01:14Preparation of Cells and the Affinity Matrix
  • 02:31Harvesting RNPs
  • 04:14Immnoprecipitation of RNP Complexes
  • 05:51Collection of Immunoprecipitated RNA
  • 07:32Results: Oligonucleotide-directed Elution of RNA-protein Complexes
  • 09:01Conclusion

Summary

Automatic Translation

Automatic Translation

This manuscript describes an approach to isolate select cognate RNPs formed in eukaryotic cells via a specific oligonucleotide-directed enrichment. We demonstrate the applicability of this approach by isolating a cognate RNP bound to the retroviral 5' untranslated region that is composed of DHX9/RNA helicase A.

Tags

Keywords: RNA ImmunoprecipitationRNA-protein ComplexesOligonucleotide-directed ElutionPost-transcriptional ControlViral InfectionsCellular DiseasesGene ExpressionMagnetic BeadsFLAG-tagged ProteinImmunoprecipitation

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