Analyses of genomic DNA isolated from livers after injection of CD-1 mice with 2 × 1011 particles of AAV-CMV-F.IX at day 1 of life. Southern blot hybridization of PCR products from liver DNA isolated from mice killed at day 5 (lanes 3 and 4) and day 240 (lanes 5 through 7) postinjection. Lane 1, no DNA added; lane 2, untransduced mouse DNA; lane 8, 0.3 copies of pAAV-CMV-F.IX per diploid mouse genome present in 50 ng of untransduced mouse DNA; lane 9, 0.06 copies of pAAV-CMV-F.IX per diploid mouse genome; mouse AP, 162-bp fragment from the mouse embryonic alkaline phosphatase gene; F-IX, 437-bp fragment from the human F.IX cDNA amplified with primer pair F.IX3/4. The average ratio of the two signals is indicated for day 5 as well as day 240 postinjection (as determined with the phosphorimaging system). Numbers in brackets are the standard deviation.
