FIG. 9.

Southern analysis of AAV rep-cap gene amplification in AAV packaging cell lines. Total cellular DNA (15 μg) was isolated from different cell lines 48 h after the indicated adenovirus infection. Episomal DNA (lane 6) was isolated from 293-GFP-145 cells after Ad-Cre infection. All DNA samples were digested with PstI to drop a 2.3-kb internal fragment of AAV cap gene. The same DNA fragment purified from AAV plasmid was used as a probe for Southern hybridized. The DNA copy number standards were set at 5, 50, and 500 rep-cap genomes per cell by spiking 0.13, 1.3, and 13 ng of plasmid pXX2-Int-3A-Hyg into 15 μg of plain 293 cell DNA. The detected band corresponds to the expected 2.3-kb cap gene fragment.