DISSERTATION
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_____________________________________________________________ Results and Discussion<br />
Figure 3.53. Scheme of the characterization sequence: a) ssDNA immobilization (1 min,<br />
0.5/-0.2 V pulse profile with 10 ms pulse duration); b) passivation with MCU (1 min, 0.5/-<br />
0.2 V pulse profile with 10 ms pulse duration); c) hybridization with Fc-tDNA (10 min<br />
incubation); d) dehybridization (H2O, 10 min); e) hybridization with a non-labelled tDNA<br />
(10 min incubation); f) intercalation with AO-GOx (15 min, incubation) and detection of<br />
the glucose oxidation current; g) dehybridization and removal of AO-GOx (H2O, ethanol,<br />
~10 min); h) evaluation of the interaction of AO-GOx with ssDNA (15 min, incubation)<br />
by amperometric detection. Chronoamperometric measurements were conducted at an<br />
applied potential of +400 mV. FAD: flavin adenine dinucleotide. All potentials vs.<br />
Ag/AgCl/3 M KCl; drawing not to scale.<br />
In order to optimize DNA detection via the synthesized AO-GOx a thorough characterization<br />
sequence was employed, which is presented in Figure 3.53. The DNA assay build-up consisted<br />
of an initial ssDNA immobilization (Figure 3.53, a) followed by passivation of the surface with<br />
MCU (Figure 3.53, b). The created DNA sensing platform was then subjected to hybridization<br />
3.5 Intercalation as a DNA detection technique 99