FIG 2.

Processivity of Dpo4 and mutants. A constant concentration (10 nM) of Cy3-labeled 36-mer/M13mp18 was added to a reaction buffer. A constant concentration of DNA polymerase (100 nM) was added to initiate DNA synthesis and a 200-fold excess of 50-mer ssDNA as a polymerase trap at 37°C for 5 min, and products formed were separated by PAGE. All reactions were done in triplicate, and the indicated data points are shown as means ± standard deviations. (A and B) The bands of products were separated by PAGE, and the A, B, and C bands are from primer extension by wtDpo4 and mutants. M, molecular mass markers. (C) The processivity value calculated by the method in combination with the length and intensity of the three main bands.