Figure 5
From: Development of a PCR Assay to detect Papillomavirus Infection in the Snow Leopard
Validation of the E6 and E7 amplification products by restriction enzyme digestion. (a) Top panel: Schematic representation of the 694-bp E6 amplification product indicating the positions of the two TaqI cleavage sites, and the sizes of the resulting fragments (288, 279 and127 bp). Bottom panel: 50 ng of genomic DNA isolated from two snow leopards (SL 1 and SL 3) using the Oragene•ANIMAL kit were amplified using the E6 primer pair. Half of each PCR reaction was incubated with the TaqI restriction endonuclease. Restriction products (lanes 2 and 4), along with an aliquot of the original PCR reactions (lanes 1 and 3) were resolved by agarose gel electrophoresis and visualized by staining with SYBR® Green. (b) Top panel: Schematic representation of 472-bp E7 amplification product indicating the positions of the three RsaI cleavage sites, and the sizes of the resulting restriction products (263, 144, 36 and 29 bp). Bottom panel: 50 ng of genomic DNA isolated from two snow leopards (SL 1 and SL 3) using the Oragene•ANIMAL kit were amplified using the E7 primer pair. Half of each PCR reaction was incubated with the TaqI restriction endonuclease. Restriction products (lanes 2 and 4), along with an aliquot of the original PCR reactions (lanes 1 and 3) were resolved by agarose gel electrophoresis and visualized by staining with SYBR® Green.