Figure 5.

ϕ29 TP-DNA amplification with wild-type and mutant TPs. The assay was carried out as described under ‘Materials and Methods’ section, in the presence of 5 ng of ϕ29 TP-DNA, 10 ng of ϕ29 DNA polymerase, 5 ng of either wild-type or the indicated mutant TP, 10 µg of ϕ29 DBP and 10 µg of ϕ29 SSB. After 80 min of incubation at 30°C, the reaction was stopped with 10 mM EDTA. The relative activity values were calculated (Table 1), and the length and amount of the synthesized DNA was analysed by alkaline agarose gel electrophoresis followed by autoradiography (upper panel) and ethidium bromide staining (lower panel).