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Visualization of SARS-CoV-2 using Immuno RNA-Fluorescence In Situ Hybridization
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JoVE 杂志 免疫与感染
Visualization of SARS-CoV-2 using Immuno RNA-Fluorescence In Situ Hybridization

Visualization of SARS-CoV-2 using Immuno RNA-Fluorescence In Situ Hybridization

DOI:
10.3791/62067-v

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05:23 min

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December 23, 2020

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, , , , , , ,
1Malopolska Centre of Biotechnology,Jagiellonian University, 2Microbiology Department Faculty of Biochemistry, Biophysics and Biotechnology,Jagiellonian University, 3Department of Cardiac, Vascular and Endovascular Surgery and Transplantology,The Medical University of Silesia in Katowice, 4Silesian Centre for Heart Diseases

Chapters

  • 00:04Introduction
  • 00:42SARS-CoV-2 RNA-FIS in Vero Cells Cultured on Coverslips
  • 02:26SARS-CoV-2 RNA-FISH in HAE Cultures
  • 03:38Results: Immuno-RNA-FISH to Detect SARS-CoV-2 RNA in Vero and HAE Cells
  • 04:38Conclusion

Summary

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Here, we describe a simple method that combines RNA fluorescence in situ hybridization (RNA-FISH) with immunofluorescence to visualize severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA. This protocol may increase understanding of the molecular characteristics of SARS-CoV-2 RNA-host interactions at a single-cell level.

Tags

Keywords: SARS-CoV-2Immuno RNA-Fluorescence In Situ HybridizationViral PathogenesisViral ReplicationSingle-cell AnalysisDiagnosticsCell LinesHuman Airway EpitheliumFixationPermeabilizationAmplification BufferHairpin ProbesSnap-coolingMounting MediumAntifadePreamplification

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