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数字液滴PCR法定量分析小鼠视网膜AAV转导效率

JoVE Journal
Neurociência
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JoVE Journal Neurociência

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

•

09:50 min

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December 25, 2021

•

Iskalen Cansu Topcu Okan², Mehri Ahmadian², Yesim Tutuncu², Halit Yusuf Altay², Cavit Agca²

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

Tadução automática

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

Keywords: Digital Droplet PCR AAV Transduction Murine Retina Quantification Absolute Quantification CRISPR Gene Therapy Mitochondrial DNA Transfection Virus Purification

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