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Propagation of Homalodisca coagulata virus-01 via Homalodisca vitripennis Cell Culture
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Immunology and Infection
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JoVE Journal Immunology and Infection
Propagation of Homalodisca coagulata virus-01 via Homalodisca vitripennis Cell Culture

Propagation of Homalodisca coagulata virus-01 via Homalodisca vitripennis Cell Culture

DOI:
10.3791/51953-v

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12:52 min

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September 25, 2014

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, , ,
1Department of Biology,University of Texas at Tyler, 2U. S. Horticultural Research Laboratory,USDA ARS

Chapters

  • 00:05Title
  • 01:52Cell Culture and Whole Virus Extraction
  • 03:56Viral Replication
  • 04:55RNA Extraction and RT-PCR
  • 07:45Confocal Microscopy
  • 09:55Results: Quantification of Cell Survivability in HoCV-01 Infected Cells
  • 12:03Conclusion

Summary

Automatic Translation

Automatic Translation

Here we present a protocol to propagate Homalodisca vitripennis cells and HoCV-1 in vitro. Medium was removed from HoCV-1 positive cultures and RNA extracted every 24 hr for 168 hr. Cell survivability was quantified by trypan blue staining. Whole virus particles were extracted post-infection. Extracted RNA was quantified by qRT-PCR.

Tags

Keywords: Homalodisca VitripennisHomalodisca Coagulata Virus-01 (HoCV-01)Xylella FastidiosaPierce’s DiseaseCell CultureViral PropagationBiopesticide ProductionQRT-PCRTrypan BlueConfocal Microscopy

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