Polymerase activity at 60°C.

<p>A) The primer extension assays used a primer labelled with FAM (fluorescein) and unlabelled template. Sites of Cy5-dCTP incorporation (Gs in the template) are shown in red. B) Primer extension time course comparing wild-type Pfu(exo-) and engineered Pfu-E10 polymerases at 60°C. Extension times are shown in minutes. Extension products used to quantify extension beyond the seven consecutive Cy5-dCTP incorporations (C₇ challenge) are highlighted in red – see Materials and Methods for details. C) Fraction of the primers extended beyond the C₇ challenge for both tested polymerases – results are shown for two independent experiments.