Fig. 3
From: A recurrent cancer-associated substitution in DNA polymerase ε produces a hyperactive enzyme
Polε-P301R has a robust DNA polymerase activity superior to that of the wild-type or proofreading-deficient Polε. a DNA polymerase activity was assayed using 25 nM correctly matched P50/T80a oligonucleotide substrate (shown above the gel image) and 6.25 nM polymerase, and the fraction of full-length product (80 nt) was quantified. Representative of two independent experiments. b Schematic of DNA replication assay on the circular M13/CAN1(1-1560-F) substrate. c M13/CAN1(1-1560-F) replication reactions were performed at a Polε:substrate ratio of 5:1. The products were separated in a 0.8% alkaline agarose gel, and the fraction of maximal-length products was quantified. Representative of two independent experiments. d The M13/CAN1(1-1560-F) replication reactions were performed at a Polε:substrate ratio of 1:5, and the products were analyzed by 8 M Urea PAGE. One experiment. Source data for a, c, and d are provided in a Source Data file
