Abstract
The development of DNA-based biosensors requires efficient immobilization of DNA probes on the sensor surfaces with optimum coverage and orientation. In this study, we have prepared thin films of a 718 base pair 5‘-thiol-labeled double-stranded DNA (dsDNA) on a gold surface by chemisorption and determined the quantity, surface coverage, and adsorption kinetics of dsDNA using cyclic voltammetry (CV), quartz crystal resonator (QCR), and radioactive labeling techniques. The adsorption and desorption rate constants of the 5‘-thiol-labeled dsDNA on a gold surface are estimated to be (1.9 ± 0.2) × 103 M-1 s-1 and (1.3 ± 1.1) × 10-3 s-1, respectively. The equilibrium constant is (1.5 ± 1.3) × 106 M-1 and the fractional coverage is >75% for the DNA concentration range of 2−10 μM. The ligand-binding properties of the immobilized dsDNA were investigated using doxorubicin, a DNA-intercalating anthracyclic antibiotic, as the probing molecule. The binding of doxorubicin to the immobilized dsDNA was examined by CV to v...
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