Preparation of electrode-immobilized, redox-modified oligonucleotides for electrochemical DNA and aptamer-based sensing
Preparation of electrode-immobilized, redox-modified oligonucleotides for electrochemical DNA and aptamer-based sensing
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DOI：
10.1038/nprot.2007.413
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Plaxco, Kevin W.
中科院分区：
文献类型：
--
作者：
Xiao, Yi;Lai, Rebecca Y.;Plaxco, Kevin W.
近年来，已经看到了许多无试剂的电化学传感器的发展，其基于靶诱导的电极结合的寡核苷酸的折叠或解折叠，迄今为止报道的实例包括用于检测特定核酸、蛋白质、小分子和无机离子的传感器。这些装置通常被称为电化学DNA（E-DNA）和E-AB（电化学，基于适体的）传感器，其由在一端用氧化还原报告物（在该方案中为亚甲蓝）修饰的寡核苷酸探针和在另一端通过硫醇-金键连接到金电极组成。分析物与寡核苷酸探针的结合改变了其结构和动力学，这反过来又影响了电子转移到询问电极的效率。这类传感器即使在直接受到血清、土壤和其他复杂的多组分样品基质的挑战时也表现良好。该方案描述了E-DNA和E-AB传感器的制造。该方案可在12 h内完成。
Recent years have seen the development of a number of reagentless, electrochemical sensors based on the target-induced folding or unfolding of electrode-bound oligonucleotides, with examples reported to date, including sensors for the detection of specific nucleic acids, proteins, small molecules and inorganic ions. These devices, which are often termed electrochemical DNA (E-DNA) and E-AB (electrochemical, aptamer-based) sensors, are comprised of an oligonucleotide probe modified with a redox reporter (in this protocol methylene blue) at one terminus and attached to a gold electrode via a thiol-gold bond at the other. Binding of an analyte to the oligonucleotide probe changes its structure and dynamics, which, in turn, influences the efficiency of electron transfer to the interrogating electrode. This class of sensors perform well even when challenged directly with blood serum, soil and other complex, multicomponent sample matrices. This protocol describes the fabrication of E-DNA and E-AB sensors. The protocol can be completed in 12 h.