Fig. 1

Illustration of the plasmids used in this study and comparison of the PCR extension rates of the DNA polymerases PfuX7, Neq2X, and Neq2X7 measured by binding of the fluorescent Pico488 dye to double-stranded DNA (A) Schematic overview of the PfuX7, Neq2X, and Neq2X7 expression plasmids. B Table with plasmids used in this study, including their Addgene IDs. C DNA polymerase activity units calculated based on incorporation of dNTPs in 10 min per mg of purified enzyme. D Fluorescence-based activity assay for PfuX7, (E) Neq2X, and (F) Neq2X7 using 0.5, 1, and 3 pmol of enzyme over time. G Fluorescence-based activity assay comparing dNTP incorporation rates between PfuX7, Neq2X, and Neq2X7 for 0.5 pmol, (H) 1 pmol, and (I) 3 pmol of each enzyme and over time. All measurements were performed in triplicates, and the graph displays the median with the standard deviation indicated. DNAP (DNA Polymerase). Pico488 measurements were done using a Synergy H1 plate reader (BioTek Instruments, Inc)