Abstract
An electrochemical DNA biosensor for the detection of NOS gene sequences from genetically modified organisms (GMOs) is presented in this paper. Single-stranded DNA (ssDNA) was covalently attached through the carboxylate ester formed by the 3′-hydroxy end of the DNA with the carboxyl of a mercaptoacetic acid self-assembled monolayer-modified gold electrode using N-hydroxysuccinimide (NHS) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC) as linkers. The electrochemical behavior of methylene blue (MB) on the ssDNA and dsDNA modified gold electrode were carefully studied. Compared with ssDNA/Au electrode, an increase of redox peak current of MB on dsDNA/Au electrode was found, which could be further used for monitoring the recognition of DNA hybridization. Based on this result, the polymerase chain reaction (PCR) product of the common inserts NOS terminator from real GMOs samples was detected successfully.
Highlights
IntroductionThe detection of specific DNA sequences is important in the fields of biochemistry and life science
The detection of specific DNA sequences is important in the fields of biochemistry and life science.Among the methods for DNA detection, an electrochemical DNA hybridization biosensor is suitable for the study of specific sequence DNA with low detection limits and high sensitivity [1,2,3,4]
The interaction of methylene blue (MB) with Single-stranded DNA (ssDNA) and dsDNA in solution was first studied by cyclic voltammetry
Summary
The detection of specific DNA sequences is important in the fields of biochemistry and life science. Among the methods for DNA detection, an electrochemical DNA hybridization biosensor is suitable for the study of specific sequence DNA with low detection limits and high sensitivity [1,2,3,4]. It is based on the immobilization of an oligonucleotide on the electrode surface and detection of the hybridization procedure with different electrochemical techniques. Millan et al [18] described a sequence-selective biosensor on self-assembled monolayer (SAM) glassy carbon electrode for DNA with Co (bpy)33+ as indicator. Schedule for the preparation of self-assembled monolayer DNA modified electrode
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