Figure 4.

Effect of primer structure on its extension by hPolε_CD. Activity of hPolε_CD in extension of a DNA primer (A), an RNA primer (B), and chimeric RNA–DNA primers with one (C), three (D), five (E), and seven (F) deoxynucleotides at the primer 3′-end was analyzed. Primer sequences are indicated on the graphs; DNA stretches in chimeric primers are underlined. Percentage of extended primer was plotted against time and the data fit to a single-exponential equation (1). Reactions, containing 0.44 µM hPolε_CD, 0.2 µM template:primer, and 1 mM dTTP, were incubated at 35 °C at the indicated time points. The mean k_obs values and standard deviations were calculated from three independent experiments using GraphPad Prizm software.