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Digital Droplet PCR Method for the Quantification of AAV Transduction Efficiency in Murine Retina
JoVE 杂志
神经科学
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JoVE 杂志 神经科学
Digital Droplet PCR Method for the Quantification of AAV Transduction Efficiency in Murine Retina

Digital Droplet PCR Method for the Quantification of AAV Transduction Efficiency in 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

Chapters

  • 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
  • 07:52Results
  • 09:07Conclusion

Summary

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This protocol presents how to quantify AAV transduction efficiency in mouse retina using digital droplet PCR (dd-PCR) together with small scale AAV production, intravitreal injection, retinal imaging, and retinal genomic DNA isolation.

Tags

Digital Droplet PCRAAV TransductionMurine RetinaQuantificationAbsolute QuantificationCRISPRGene TherapyMitochondrial DNATransfectionVirus Purification

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