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Semi-quantitative Detection of RNA-dependent RNA Polymerase Activity of Human Telomerase Reverse Transcriptase Protein
JoVE Journal
Genetics
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JoVE Journal Genetics
Semi-quantitative Detection of RNA-dependent RNA Polymerase Activity of Human Telomerase Reverse Transcriptase Protein

Semi-quantitative Detection of RNA-dependent RNA Polymerase Activity of Human Telomerase Reverse Transcriptase Protein

DOI:
10.3791/57021-v

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08:26 min

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June 12, 2018

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, , , ,
1Division of Cancer Stem Cell,National Cancer Center Research Institute

Chapters

  • 00:04Title
  • 00:29Immunoprecipitation (IP)-RdRP Assay: Preparation of Cell Lysate
  • 01:34IP-RdRP Assay: Immunoprecipitation
  • 03:54IP-RdRP Assay: RdRP Reaction
  • 06:19IP-RdRP Assay: RNase Treatment
  • 07:38Results: RdRP Products of Endogenous TERT in Mitotic HeLa Cells
  • 08:10Conclusion

Summary

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Human telomerase reverse transcriptase (TERT) synthesizes not only telomeric DNA but also double-stranded RNA through RNA-dependent RNA polymerase activity. Here, we describe a newly established assay to detect RNA-dependent RNA polymerase activity of endogenous TERT.

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Keywords: RNA-dependent RNA PolymeraseTERTRNA BiologyRdRP ActivityHeLa CellsCell LysisImmunoprecipitationProtein PurificationMicrococcal NucleaseSemi-quantitative Detection

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