Voltammetric determination of DNA hybridization using methylene blue and self-assembled alkanethiol monolayer on gold electrodes

被引:247
作者
Kerman, K
Ozkan, D
Kara, P
Meric, B
Gooding, JJ
Ozsoz, M [1 ]
机构
[1] Ege Univ, Fac Pharm, Dept Analyt Chem, TR-35100 Izmir, Turkey
[2] Univ New S Wales, Sch Chem Sci, Sydney, NSW 2052, Australia
关键词
DNA biosensor; DNA; methylene blue; hybridization; self-assembled monolayers; inosine;
D O I
10.1016/S0003-2670(02)00308-2
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
An electrochemical DNA biosensor based on the recognition of single stranded DNA (ssDNA) by hybridization detection with immobilized complementary DNA oligonucleotides is presented. DNA and oligonucleotides were covalently attached through free amines on the DNA bases using N-hydroxysulfosuccinimide (NHS) and N-(3-dimethylamino)propyl-N'-ethylcarbodiimide hydrochloride (EDC) onto a carboxylate terminated alkanethiol self-assembled monolayers (SAM) preformed on a gold electrode (AuE). Differential pulse voltammetry (DPV) was used to investigate the surface coverage and molecular orientation of the immobilized DNA molecules. The covalently immobilized probe could selectively hybridize with the target DNA to form a hybrid on the surface despite the bases being attached to the SAM. The changes in the peak currents of methylene blue (MB), an electroactive label, were observed upon hybridization of probe with the target. Peak currents were found to increase in the following order: hybrid-modified AuE, mismatched hybrid-modified AuE, and the probe-modified AuE which indicates the MB signal is determined by the extent of exposed bases. Control experiments were performed using a non-complementary DNA sequence. The effect of the DNA target concentration on the hybridization signal was also studied. The interaction of MB with inosine substituted probes was investigated. Performance characteristics of the sensor are described. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:39 / 47
页数:9
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