Electrochemical DNA quantification based on aggregation induced by Hoechst 33258

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Abstract A novel DNA quantification method by using a redox-active molecule, Hoechst 33258, 2 ′ -(4-hydroxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5 ′ -bi(1H-benzimidazole), is reported. Hoechst 33258 was interacted with DNA in solution without immobilization on the electrode surface, thus the time-consuming probe immobilization step was eliminated. The changes in the anodic current signal of Hoechst 33258 at 0.50 V was monitored in the presence and absence of solution-phase DNA by using a bare glassy carbon electrode (GCE) in connection with linear sweep voltammetry (LSV). The interaction of DNA with several small molecules was also monitored by using cyclic voltammetry (CV) at a bare GCE in order to determine the most effective molecule for DNA aggregation. Hoechst 33258 was found to form an aggregate in the presence of DNA, and this phenomenon was confirmed by using atomic force microscopy (AFM). The aggregation of DNA–Hoechst 33258 complex led to a decrease in the voltammetric signal in proportion to the quantity of DNA in a wide linear range between 1.5 and 25 mg/l. The interaction of Hoechst 33258 with polynucleotides was also monitored to determine its sequence specificity. DNA amplification by polymerase chain reaction (PCR), and the detection of DNA sequences related to Salmonella enteritidis , Streptococcus sobrinus , and hepatitis B virus (HBV) were demonstrated by using DNA- Hoechst 33258 aggregation system.

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