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. 2013 Aug 7;24(9):814–819. doi: 10.1089/hum.2013.118

FIG. 2.

FIG. 2.

FIG. 2.

Detection of Ad sequences in cellular DNA from liver tissue of a patient who received AdOTC gene transfer. Using optimized primers and PCR conditions, 200 ng each of cellular DNA from liver tumor (A, C, D) and normal tissue (B, C, E) of an OTC deficiency heterozygote who received AdOTC gene transfer more than 10 years ago were subjected to nested PCR for amplifications of Ad5 DNA polymerase gene (A, B), hOTC transgene/remnant E1 gene junction (D, E), and native E1 gene (C) sequences. PCR amplification of Ad sequence in 200 ng each of cellular DNA from naive mouse liver tissue spiked with 1 GC of the isolated rH5.001CB-hOTC vector DNA (A, B, D, E) and wtAd plasmid (pH5wt) DNA (C) were used as positive controls. PCR products were detected by 1.5% agarose gel electrophoresis and verified by TOPO-cloning followed by sequencing. T, tumor; LL, left adjacent normal liver; RL, right adjacent normal liver; mock, no DNA template control; M, molecular weight ladder.