Adenovirus Production: How to Clone, Package, and Harvest Adenovirus for Your Seed Stocks

[@abmgood]

Here are some helpful time stamps for our adenovirus packaging video: 1:50 How to clone your gene of interest into an adenovirus expression system. 3:07 How to amplify and linearize your recombinant adenovector for use in the transfection step. 3:41 Preparing your HEK293 cells for transfection 4:21 How to prepare your transfection solutions. 5:45 How to transfect your HEK293 cells. 6:15 Checking your cells for successful transfection 6:32 How to make your adenovirus seed stock and amplify it to achieve a higher titer! 7:00 How to harvest and store your adenoviruses.

[@samiurrahmansefat1336]

Why do we linearize adenoviral vector before transfection? The obvious answer might be that it is tough to deliver a large plasmid into cells. Are there any additional reasons?

[@maxbailey9164]

Think you’ve hit the nail on the head. From what I understand, transfection is relatively poorly understood, but I’d imagine it’s similar to the linearisation of a plasmid before being run on a gel