Fig. 5.

Assessing the effects of repetitive RCA on resulting viral titer. We examined the impact of sequential RCA on the ability to rescue virus in Vero cells. We used the Evomics SuperPhi RCA kit and a sample of midiprepped DNA as a template to generate RCA products (Passage 0). We then used the RCA product as the template for subsequent RCA reactions (Passage 1–3). We transfected the cells using 500 ng of RCA product or 500 ng of plasmid DNA in triplicate. The supernatant was collected 2-days post-transfection for plaque assay. Error bars represent the standard deviation from the mean. Statistical analysis was performed using one-way ANOVA with ad hoc Dunnett’s correction for multiple comparisons against the plasmid control (ns P > 0.05, **P ≤ 0.01, ***P ≤ 0.001).