1Biotechnology Division, Department of Molecular and Cell Biology, Faculty of Chemistry, University of Kashan, Kashan, Iran
2Department of Analytical Chemistry, Faculty of Chemistry, University of Kashan, Kashan, Iran
Over the past years, DNA biosensors have been developed to analyze DNA interaction and damage that have important applications in biotechnological researches. The immobilization of DNA onto a substrate is one key step for construction of DNA electrochemical biosensors. In this report, a direct approach has been described for immobilization of single strand DNA onto carboxylic acid-functionalized carbon nanotubes modified glassy carbon electrode. To do this, we first modified the glassy carbon electrode surface with MWCNT-COOH. The immersion of MWCNT-COOH/GCE in ss-DNA probe solution, with different pH and ionic strength, was followed by suitable interaction between amine group of ss-DNA bases and carboxylic groups of MWCNT-COOH. This interaction leads to successful ss-DNA immobilization on MWCNT-COOH that was confirmed by cyclic voltammetry, electrochemical impedance spectroscopy and atomic force microscopy. Immobilization of ss-DNA on the modified electrode increased the charge transfer resistant but decreased the peak current of redox probe ([Fe(CN)6]3-/4-). The result of cyclic voltammograms implicates that enhancements in the DNA immobilization are possible by adroit choice of low pH and high ionic strength. The standard free-energy of adsorption (ΔG°ads) was calculated from electrochemical impedance spectroscopy data (-47.75 kJ mol-1) and was confirmed covalent bond formation. atomic force microscopy topographic images demonstrate increased surface roughness after ss-DNA immobilization. Results offer a simple, rapid and low-cost of DNA immobilization strategy can be opportunities to design of novel nucleic acid biosensors.
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