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Investigation of RNA Synthesis Using 5-Bromouridine Labelling and Immunoprecipitation
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JoVE Revista Genética
Investigation of RNA Synthesis Using 5-Bromouridine Labelling and Immunoprecipitation

Investigation of RNA Synthesis Using 5-Bromouridine Labelling and Immunoprecipitation

DOI:
10.3791/57056-v

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09:59 min

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May 03, 2018

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, , , ,
1DANDRITE – Danish Research Institute of Translational Neuroscience, Dept. of Biomedicine,Aarhus University, 2Dept. of Molecular Biology and Genetics,Aarhus University

Capítulos

  • 00:04Título
  • 00:37RNA Sample Preparation
  • 02:37Immunoprecipitation Bead Preparation
  • 04:51Binding BrU-labelled RNA to Beads
  • 05:46RNA Elution Purification
  • 08:29Results: Increase in Steady State RNA Caused by Increase in RNA Synthesis
  • 09:35Conclusion

Summary

Traducción Automática

Traducción Automática

This method can be used to measure RNA synthesis. 5-Bromouridine is added to cells and incorporated into synthesized RNA. RNA synthesis is measured by RNA extraction immediately after labelling, followed by 5-Bromouridine-targeted immunoprecipitation of labelled RNA and analysis by reverse transcription and quantitative polymerase chain reaction.

Tags

RNA Synthesis5-bromouridine LabelingImmunoprecipitationHEK293T CellsRNA ExtractionMagnetic BeadsBromouridine-IP Buffer

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