Adenovirus culture/isolation - (Aug/16/2005 )

Hello all-

I am starting new work growing adenovirus, which we will isolate & then run PCR. I have not worked with viral cultures before (background is parasitology!), & I need to figure out what is the best way to isolate the virus; I was told that it will grow relatively easy, the problem is isolating the DNA. Any ideas would be much appreciated for this very novice virologist!!!!

Thanks!!

Hi

Adenovirus will grow well in HEK293 cells, as these have integrated viral genome, and hence will allow packaging of the viral particles.

To isolate the DNA, isolate the virus using CsCl gradient or some other method (like adenopack from vivascience) if you can afford it. Once you have the virus you can lyse them with proteinase K and extract the DNA from there.

Bob

thanks so much for the reply. if you get the reagents etc. necessary to perform the CsCl procedure, would that likely be the same amount of $$ as the adenopack kit? have you tried the kit, does it give as high yield as it claims?

thanks again-

Also...would anyone happen to have a good CsCl procedure??

Thanks!