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Måling av BK-polyomavirus ikke-Coding Control region drevet Transcriptional aktivitet via Flow flowcytometri
JoVE Journal
Genetics
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JoVE Journal Genetics
Measurement of BK-polyomavirus Non-Coding Control Region Driven Transcriptional Activity Via Flow Cytometry
DOI:

11:54 min

July 13, 2019

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Chapters

  • 00:00Title
  • 01:15Collection of Blood and or Urine Samples and Isolation of BKPyV-DNA
  • 02:11Amplification and Sequencing of the Non-coding Control Region (NCCR)
  • 03:37Cloning of the Non-coding Control Region (NCCR) into the Dual Fluorescence Reporter
  • 05:26Transient Transfection of HEK293T Cells with the Dual Fluorescence Reporter Plasmid and Treatment with Potential Antiviral Agents
  • 07:23Fluorescence Microscopy and Flow Cytometry
  • 09:22Results and Data Interpretation
  • 11:26Conclusion

Summary

Automatic Translation

I dette manuskriptet presenteres en protokoll for å utføre FACS-basert måling av BK-polyomavirus transcriptional aktivitet ved å bruke HEK293T celler transfekterte med en toveis reporter plasmider som uttrykker tdTomato og eGFP. Denne metoden videre gjør det mulig å kvantitativt bestemme innflytelsen av romanen forbindelser på viral transkripsjon.

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