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吸附转移溶出伏安法:测定固定在电极表面的纳克量DNA。

Adsorptive transfer stripping voltammetry: determination of nanogram quantities of DNA immobilized at the electrode surface.

作者信息

Palecek E

机构信息

Institute of Biophysics, Czechoslovak Academy of Sciences, Brno.

出版信息

Anal Biochem. 1988 May 1;170(2):421-31. doi: 10.1016/0003-2697(88)90654-9.

Abstract

In adsorptive transfer stripping voltammetry (AdTSV), DNA is first adsorbed at the electrode, the electrode is washed and transferred (with the adsorbed layer) in the medium not containing DNA, and voltammetric analysis is performed in this medium. Adsorption can be performed from a drop of DNA solution, which makes it possible to reduce the volume of the analyzed sample by two orders of magnitude as compared to that of conventional voltammetry. With the hanging mercury drop electrode the limit of detection of single-stranded DNA is below 0.1 micrograms/ml; thus if the adsorption is performed from a 10-microliter drop of DNA solution subnanogram quantities of single-stranded DNA are sufficient for the analysis. In AdTSV the behavior of single- and double-stranded DNAs markedly differ from each other in a manner similar to that in the conventional voltammetric or polarographic analysis; AdTSV can thus be used in DNA structure analysis. In AdTSV the DNA transport and its adsorption at the electrode are separated from the electrode process; due to this fact it is possible (a) to perform the voltammetric analysis of DNA from media not suitable for voltammetric analysis of the conventional type, (b) to study the interaction of immobilized DNA with other substances in solution without the results of the voltammetric analysis being influenced by DNA interactions in the bulk of solution, and (c) to exploit the differences of adsorbability of DNA and other substances in order to separate them on the electrode.

摘要

在吸附转移溶出伏安法(AdTSV)中,DNA首先吸附在电极上,然后将电极冲洗并(连同吸附层一起)转移到不含DNA的介质中,在此介质中进行伏安分析。吸附可以从一滴DNA溶液中进行,这使得与传统伏安法相比,分析样品的体积可以减少两个数量级。使用悬汞滴电极时,单链DNA的检测限低于0.1微克/毫升;因此,如果从10微升的DNA溶液滴中进行吸附,亚纳克量的单链DNA就足以进行分析。在AdTSV中,单链和双链DNA的行为在方式上与传统伏安法或极谱分析中的行为明显不同;因此,AdTSV可用于DNA结构分析。在AdTSV中,DNA在电极上的传输及其吸附与电极过程是分开的;由于这一事实,有可能(a)从不适用于传统类型伏安分析的介质中对DNA进行伏安分析,(b)研究固定化DNA与溶液中其他物质的相互作用,而伏安分析结果不受溶液主体中DNA相互作用的影响,以及(c)利用DNA和其他物质吸附性的差异在电极上对它们进行分离。

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