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A Quantitative Dot Blot Assay for AAV Titration and Its Use for Functional Assessment of the Adeno-associated Virus Assembly-activating Proteins
JoVE Journal
Biology
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JoVE Journal Biology
A Quantitative Dot Blot Assay for AAV Titration and Its Use for Functional Assessment of the Adeno-associated Virus Assembly-activating Proteins
DOI:

14:49 min

June 12, 2018

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Chapters

  • 00:04Title
  • 00:46Production of AAV in HEK 293 Cells by Plasmid DNA Transfection
  • 02:50Recovery of Viral Particles, and Treatment with Serratia marcescens Endonuclease and Proteinase K
  • 04:20Phenol-choloroform Extraction, Ethanol Precipitation, and Preparation of Plasmid DNA Standards
  • 05:55Dot Blot Assay
  • 08:46Hybridization and Washing
  • 10:50Results: AAV Titration and Functional Assessment of AAP Proteins Using a Quantitative Dot Blot Assay
  • 12:58Conclusion

Summary

Automatic Translation

This manuscript details a straightforward dot blot assay for quantitation of adeno-associated virus (AAV) titers and its application to study the role of assembly-activating proteins (AAPs), a novel class of non-structural viral proteins found in all AAV serotypes, in promoting the assembly of capsids derived from cognate and heterologous AAV serotypes.

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