PCR Plate Spinner, 120V

Company: VWR

Catalog#: 89184-608

Quantitation of Regulatory Activity for the Complement Alternative Pathway Using an Adaptation of the AP₅₀ in vitro Assay

Sheila Cabezas Falcon, David L Gordon and Jillian M Carr

Published online: 2018-12-20

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Virus Binding and Internalization Assay for Adeno-associated Virus

Author:

Garrett E. Berry and Longping V. Tse,

Date:

2017-01-20

[Abstract] The binding and internalization of adeno-associated virus (AAV) is an important determinant of viral infectivity and tropism. The ability to dissect these two tightly connected cellular processes would allow better understanding and provide insight on virus entry and trafficking. In the following protocol, we describe a quantitative PCR (qPCR) based method to determine the amount of vector bound to the cell surface and the amount of subsequent virus internalization based on viral genome quantification. This protocol is optimized for studying AAV. Nevertheless, it can serve as a backbone for studying other viruses with careful modification. [摘要] 腺相关病毒（AAV）的结合和内化是病毒感染和向性的重要决定因素。解决这两个紧密相连的细胞过程的能力将有助于更好地了解并提供有关病毒进入和贩运的洞察。在以下协议中，我们描述了基于定量PCR（qPCR）的方法，以确定基于病毒基因组定量结合到细胞表面的载体的量和随后的病毒内化的量。该协议针对研究AAV进行了优化。然而，它可以作为研究其他病毒，仔细修改的骨干。
【背景】评估AAV生物学的研究通常使用转基因表达作为实验终点。然而，AAV在到达细胞核并转导细胞之前必须成功导航的许多关键步骤。因此，AAV感染途径中有多个不同的步骤可能会单独或共同破坏，导致改变的转导。 AAV结合和内化的评估是确定小分子细胞修饰，基于CRISPR的基因敲除，基于siRNA的基因敲除或其他实验程序后观察到的转导差异的原因的重要的第一步

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PCR Plate Spinner, 120V

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