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A Microfluidic-based Electrochemical Biochip for Label-free DNA Hybridization Analysis
Published on: September 10, 2014
Differential pulse voltammetric studies of ethidium bromide binding to DNA
S H Minasyan1, L A Tavadyan, A P Antonyan
1Laboratory of Liquid-Phase Oxidation and Free-Radical Reactions, Institute of Chemical Physics named after A.B. Nalbandyan, NAS of Armenia, Sevak St., 5/2, 375014, Yerevan, Armenia.
Abstract:
The interaction of ethidium bromide (EtBr) with calf thymus DNA is investigated electrochemically with the use of differential pulse voltammetry (DPV) at two different ionic strengths of a solution (0.154 M and 0.02 M [Na+], pH 7.0). It is revealed that EtBr binds with DNA in more than one way. The appropriate values of constants (K) and number site sizes (n) of EtBr binding to DNA are determined. The values of binding constants are equal to 1.9 x 10(6) and 5.6 x 10(5) M(-1), and number site sizes to 9 and 3.6 for strong interactions at ionic strengths of solutions 0.02 and 0.154 M Na+ at 28 degrees C, respectively. For a weaker interaction, these parameters are equal to 7 x 10(4) and 8 x 10(4) M(-1) and 1.5 and 1 at the mentioned ionic strengths of solutions, respectively. Thus, EtBr interacts with DNA in more than one way--intercalative and electrostatic at low ionic strength, and semi-intercalative and electrostatic at a higher strength of the solution. These results are in good accordance with the ones obtained by spectroscopic (absorption and fluorimetric) methods.
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