AAV plasmids containing different versions of the G protein gene were submitted to the Viral Vector Core of The Research Institute at Nationwide Children’s Hospital (Columbus, Ohio, USA) for AAV production as described [36]. Briefly, recombinant self-complementary AAV vector plasmids containing the genes of interest were transfected into adherent HEK293 cells with AAV serotype 5 and Ad helper plasmids by CaPO4 transfection. Cell were fragmented and AAV particles were purified by iodixanol step gradient ultracentrifugation and ion exchange column chromatography [40,41]. AAV titers were determined by qPCR and purity was confirmed by SDS-PAGE as previously described [40,41].

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.