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Digital droplet PCR-metode til kvantificering af AAV Transduktionseffektivitet i Murine Retina
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JoVE Journal Neurociência
Digital Droplet PCR Method for the Quantification of AAV Transduction Efficiency in Murine Retina

Digital droplet PCR-metode til kvantificering af AAV Transduktionseffektivitet i Murine Retina

DOI:
10.3791/63038-v

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09:50 min

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December 25, 2021

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, , , ,
1Molecular Biology, Genetics and Bioengineering Program,Sabanci University, 2Nanotechnology Research and Application Center (SUNUM),Sabanci University

Capítulos

  • 00:01Introduction
  • 00:57Small Scale AAV Production
  • 02:52Intravitreal Injection of AAV
  • 04:17Fluorescence and Fundus Imaging
  • 04:48Retina Isolation
  • 05:16Tissue Genomic DNA Isolation
  • 05:57Digital Droplet PCR Analysis of Mouse Retina Samples for Quantification of Viral Genomes
  • 07:52Resultados
  • 09:07Conclusion

Summary

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Tadução automática

Denne protokol præsenterer, hvordan man kvantificerer AAV transduktion effektivitet i musen nethinden ved hjælp af digital dråbe PCR (dd-PCR) sammen med lille skala AAV produktion, intravitreal injektion, retinal billeddannelse, og retinal genomisk DNA isolation.

Tags

Keywords: Digital Droplet PCRAAV TransductionMurine RetinaQuantificationAbsolute QuantificationCRISPRGene TherapyMitochondrial DNATransfectionVirus Purification

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