Addgene Protocols Go Viral: AAV Purification and Titration

By Eddy Page

Since rolling out our viral service in 2016, Addgene has shipped thousands of AAV and lentiviral preps to scientists around the world. In the years since, our scientists have optimized many steps in the viral production and quality control process to improve both the yield and quality of the viral preps. For example, last year we published an article describing a sucrose supplementation strategy to improve yield. In addition, as described in this blog post, the team developed a viral genome sequencing strategy to confirm the identity and purity of all AAV viral lots.

Aligning with Addgene’s mission to “accelerate research and discovery by improving access to useful research materials and information,” we love to share lab protocols that we use as we serve the scientific community. Here are a few virus-related protocol videos that may help to save you time and improve your experiment results!

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AAV purification by iodixanol gradient ultracentrifugation

This video goes over AAV vector purification using an iodixanol column gradient. We'll show the loading and unloading of the column and give some tips for performing this purification.  Check out the AAV Purification protocol page for more detailed instructions.

AAV titration by qPCR using SYBR green technology

Here’s some advice on how to successfully set up your qPCR to determine the genome copies in a given AAV preparation. For detailed instructions and calculations on how to prepare a plasmid stock used to generate a standard curve, visit the AAV Titration by qPCR written protocol page.

Getting started with tissue culture

The cultivation of mammalian cells in the lab, or tissue culture as it is commonly called, is a critical tool for many scientists. Mammalian cells provide scientists with the means to study biological processes on the cellular level instead of having to work with a whole organism and can help scientists produce vital tools in the lab such as antibodies or viruses. If you’re new to tissue culture, check out this video for some tips and best practices for getting started with tissue cultures.

To keep up on all of the latest protocol videos, lab tips, and more video content from Addgene, subscribe to Addgene’s YouTube Channel.

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Topics: Viral Vectors, Viral Vector Protocols and Tips, AAV

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