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一种将非标记单链 DNA 静电固定在自组装重氮修饰金电极上的新方法及其杂交的差分脉冲伏安法检测。

A new methodology for electrostatic immobilization of a non-labeled single strand DNA onto a self-assembled diazonium modified gold electrode and detection of its hybridization by differential pulse voltammetry.

机构信息

Faculty of Chemistry, Kharazmi (Tarbiat Moallem) University, Tehran, Iran.

出版信息

Talanta. 2013 Jan 15;103:344-8. doi: 10.1016/j.talanta.2012.10.069. Epub 2012 Oct 29.

DOI:10.1016/j.talanta.2012.10.069
PMID:23200397
Abstract

In this work, a self-assembled diazonium modified gold electrode (D-MGE) was used for the fabrication of an electrochemical DNA biosensor. For preparation of D-MGE, initially 2-amino-5-mercapto-1, 3, 4-thiadiazole (AMT) self-assembled on gold electrode and a simple diazonation reaction was used to prepare D-MGE. Then, non-labeled single strand DNA (NL-ssDNA) directly was immobilized on D-MGE through electrostatic interaction. Cyclic voltammetry and electrochemical impedance spectroscopy (EIS) characterized the DNA biosensor fabrication process with the use of ferro-ferric cyanide as an electrochemical redox indicator. The hybridization capacity of the developed biosensor was monitored with differential pulse voltammetry using Fe (CN)(6) as an indicating probe. A wide dynamic detection range (7.9 × 10(-11)-1.2 × 10(-7) mol L(-1)) and a low detection limit (1.4 × 10(-11) mol L(-1)) were achieved for the complementary sequence. In addition, the hybridization specificity experiments showed that the sensing system could accurately discriminate complementary sequence from non-complementary sequences.

摘要

在这项工作中,使用自组装的重氮修饰金电极(D-MGE)来制备电化学 DNA 生物传感器。为了制备 D-MGE,首先将 2-氨基-5-巯基-1,3,4-噻二唑(AMT)自组装在金电极上,并通过简单的重氮化反应制备 D-MGE。然后,非标记的单链 DNA(NL-ssDNA)通过静电相互作用直接固定在 D-MGE 上。使用亚铁氰化铁作为电化学氧化还原指示剂,循环伏安法和电化学阻抗谱(EIS)对 DNA 生物传感器的制备过程进行了表征。使用 [Fe(CN)(6)] (4-)作为指示探针,通过差分脉冲伏安法监测开发的生物传感器的杂交能力。对于互补序列,实现了宽的动态检测范围(7.9×10(-11)-1.2×10(-7) mol L(-1))和低检测限(1.4×10(-11) mol L(-1))。此外,杂交特异性实验表明,传感系统能够准确地区分互补序列和非互补序列。

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