Ad5/mT and Ad5/Gp virus strains are only able to replicate in and lyse 293 N3S cells,
because the cells contain integrated copies of the E1A region.Therefore, to produce
virus stock grow cells to 80% of confluency and infect the cells with either Ad5/mT or
Ad5/Gp virus-vector (see above). Incubate cell dishes for at least two days in the virus
tissue culture cell incubator. After that remove media (cells and debris) and centrifuge
in the ICE clinical centrifuge (in tissue culture) , 3000 rpm at room temperature (RT) for
5 min. Remove supernatant and freeze it at -80ºC. To determine the pfu/ml (=titer) of
your stock a plaque assay has to be carried out.
