GENERATION AND IDENTIFICATION OF RECOMBINANT ADENOVIRUS BY LIPOSOME-MEDIATED TRANSFECTION AND PCR ANALYSIS

A simplified method for generating recombinant adenovirus was develope d by using liposome-mediated co-transfection and by directly observing for the cytopathic effect caused by recombinant adenovirus in transfe cted 293 cells. This approach avoided difficulties associated with cal cium-phosphate precipitation and agarose overlays for plaque assays. T he ease of generating recombinant adenovirus was considerably improved . Analysis by PCR of DNA samples from the supernatant of the cell cult ures with the cytopathic effect was also developed, which made identif ication of any newly generated recombinant virus rapid and specific.