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. 2015 Jul 15;5:12066. doi: 10.1038/srep12066

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Copyright © 2015, Macmillan Publishers Limited

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A: Schematic of the ss-dsDNA which is converted to dsDNA during dNMP incorporation. As a result, the polymerase moves outward along the DNA nanolever (which increases the friction and slows the DNA switching) and the dye is being held at a greater distance from the surface by the rigid double helix when the DNA is standing (which increases the fluorescence). B: Taq was bound to ss-dsDNA and baselines are recorded for 1.5 min (no dissociation). The injection of a dNTP-mix (100 μM) leads to a rapid incorporation of nucleotides (DR_up-drop and F_up-rise), followed by the dissociation of polymerase from the dsDNA end (increasing DR_up and slightly decreasing F_up). C: Temperature-dependent real-time elongation measurements (c_dNTP = 100 μM). Lines are single exponential fits ∝ exp{−t}. D: Elongation rates from C as a function of temperature, the line is an Arrhenius fit.

Inline graphic — A: Schematic of the ss-dsDNA which is converted to dsDNA during dNMP incorporation. As a result, the polymerase moves outward along the DNA nanolever (which increases the friction and slows the DNA switching) and the dye is being held at a greater distance from the surface by the rigid double helix when the DNA is standing (which increases the fluorescence). B: Taq was bound to ss-dsDNA and baselines are recorded for 1.5 min (no dissociation). The injection of a dNTP-mix (100 μM) leads to a rapid incorporation of nucleotides (DR_up-drop and F_up-rise), followed by the dissociation of polymerase from the dsDNA end (increasing DR_up and slightly decreasing F_up). C: Temperature-dependent real-time elongation measurements (c_dNTP = 100 μM). Lines are single exponential fits ∝ exp{−t}. D: Elongation rates from C as a function of temperature, the line is an Arrhenius fit.