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. 2010 Jun 10;7:123. doi: 10.1186/1743-422X-7-123

Figure 1.

Figure 1

Transient replication of HPV16 DNA in different cell types. Five μg of plasmid containing HPV16 genomic DNA were transfected into various cell lines; baby hamster kidney cell (BHK), American green monkey kidney cell (Vero), human osteoblastoma cell (U2OS), human cervical carcinoma cell (C-33A) and human embryonic kidney cell (293). After 4 days of transfection, low molecular weight DNA was extracted from 1 × 107 cells by Hirt method. During extraction, 10 ng of pUC19 DNA was added as a "spiked plasmid" to monitor the recovery of the plasmid DNA from transfected cells and to test the completeness of DpnI digestion. For the Southern analysis, 10% of Hirt-DNA was digested with HindIII to linearize the plasmid and 90% was digested with both HindIII and DpnI. The blot was probed with random-primed32-P labeled pUC19 DNA. As a control, linearized plasmid containing HPV16 at concentrations of 100 and 10 pg was loaded on the left-most lanes of the blot.