3. Once completed, datasets generated during the “Kinetics” step
are loaded in the switchANALYSIS software package. For each
concentration of polymerase, three distinct kinetics could be
successively analyzed: k ON , k POL , and k OFF . If several concentrations of polymerase have been used, a global fit can be used
in order to significantly improve the accuracy of each fitted
value. Polymerization rates (in residue s
À1 ) are calculated considering a 44 base elongated DNA.
Fig. 2 Example of DNA polymerization experiment using four HIV-1 RT concentrations. Fluorescence signal
observed upon addition of HIV-1 RT onto DNA nanolevers (a). on- (b) and off (c) rates obtained using the
switchANALYSIS software package. Determination of the k pol using switchANALYSIS
150
Guillaume Bec and Eric Ennifar
are loaded in the switchANALYSIS software package. For each
concentration of polymerase, three distinct kinetics could be
successively analyzed: k ON , k POL , and k OFF . If several concentrations of polymerase have been used, a global fit can be used
in order to significantly improve the accuracy of each fitted
value. Polymerization rates (in residue s
À1 ) are calculated considering a 44 base elongated DNA.
Fig. 2 Example of DNA polymerization experiment using four HIV-1 RT concentrations. Fluorescence signal
observed upon addition of HIV-1 RT onto DNA nanolevers (a). on- (b) and off (c) rates obtained using the
switchANALYSIS software package. Determination of the k pol using switchANALYSIS
150
Guillaume Bec and Eric Ennifar