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数字液滴PCR法定量分析小鼠视网膜AAV转导效率
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Neuroscience
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JoVE Journal Neuroscience
Digital Droplet PCR Method for the Quantification of AAV Transduction Efficiency in Murine Retina
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数字液滴PCR法定量分析小鼠视网膜AAV转导效率

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

Automatic Translation

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该协议介绍了如何使用数字液滴PCR（dd-PCR）以及小规模AAV生产，玻璃体内注射，视网膜成像和视网膜基因组DNA分离来量化小鼠视网膜中的AAV转导效率。

Tags

Digital Droplet PCRAAV TransductionMurine RetinaQuantificationAbsolute QuantificationCRISPRGene TherapyMitochondrial DNATransfectionVirus Purification

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