Electrochemical Detection of Klenow Fragment Exo-(KF-) Activity Based on Gold Nanoparticles Amplification

doi:10.3969/j.issn.0251-0790.2012.09.012

Abstract

Abstract:

A label-free, simple and fast electrochemical method for Klenow fragment exo-(KF^-) detection was developed based on the properties of gold nanoparticles(AuNPs) such as the large specific surface, small size and the ability of binding much DNA. In this work, thiolated primer DNA was first modified on the surface of electrode. In the presence of template DNA and reporter DNA tethered with AuNPs(AuNPs-DNA), primer DNA and report DNA were hybridized with template DNA to form a sandwich-like hybridization mode, which brings AuNPs-DNA to the electrode surface. When put the modified electrode into the electrolyte containing [Ru(NH₃)₆]³⁺(RuHex), the RuHex quantitatively binds to the DNA via electrostatic interaction. Since AuNPs was loaded with much reporter DNA that can bind a large number of RuHex, this offers the electrochemical signals amplification for the detection. In the presence of dNTPs and KF^-, the extension reaction was triggered, which results in the dissociation of the AuNPs-DNA and release of abundant RuHex molecules. As a result, the electrochemical signal was reduced and the KF^- was detected. Using this method, a detection limit as low as 5 U/mL for KF^- was achieved.

Key words: Klenow fragment exo-, Gold nanoparticles, [Ru(NH₃)₆]³⁺, Signal amplification

CLC Number:

O657.1

TrendMD:

LIU Jin-Quan, NI Xiao-Qi, HE Xiao-Xiao, WANG Yong-Hong, WANG Ke-Min, CHEN Zhi-Feng, SU Jing, YAN Gen-Ping. Electrochemical Detection of Klenow Fragment Exo-(KF^-) Activity Based on Gold Nanoparticles Amplification[J]. Chem. J. Chinese Universities, 2012, 33(09): 1945.

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Electrochemical Detection of Klenow Fragment Exo-(KF^-) Activity Based on Gold Nanoparticles Amplification

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