Figure 5.

Use of wild-type and chimerical TPs by the chimerical DNA polymerase with ssDNA as template. The TP-dAMP formation assay was carried out as described in Materials and Methods section, in the presence of 1 mM MnCl₂, and using 300 nM of either wild-type or chimerical DNA polymerase, 600 nM of the indicated TP and 5.6 µM of ssDNA LOT12 as template. After incubation for 10 min at 30°C, the reactions were stopped, processed, and analysed by SDS–PAGE and autoradiography. The position of TP-dAMP is indicated. For clarity, both ϕ29 wild-type and chimerical DNA polymerase are depicted on top of the figure, following the same colour code as in Figure 2.