Publication details

Voltammetric analysis of 5-(4-Azidophenyl)-2 '-deoxycytidine nucleoside and azidophenyl-labelled single- and double-stranded DNAs

Authors

DANHEL Ales TROSANOVA Zuzana BALINTOVA Jana HAVRAN Ludek HOCEK Michal BAREK Jiri FOJTA Miroslav

Year of publication 2016
Type Article in Periodical
Magazine / Source Electrochimica Acta
MU Faculty or unit

Central European Institute of Technology

Citation
Web http://www.sciencedirect.com/science/article/pii/S0013468616318205
Doi http://dx.doi.org/10.1016/j.electacta.2016.08.096
Field Electrochemistry
Keywords Aromatic Azide; Enzymatic Incorporation; Mercury Electrode; Nucleic Acid; Redox Labeling; Voltammetry
Description Voltammetric determination of a redox labeled nucleoside 5-(4-azidophenyl)-2'-deoxycytidine (dC(AZP)) and various polymerase-synthesized dC(AZP)-labeled DNAs in aqueous buffers is presented. Influence of: i) pH (2-12), ii) scan rates (0.02-10 V s(-1)), and iii) dC(AZP) concentration (0.02-10 mu mol l(-1)), on voltammograms of dC(AZP) were systematically studied for the first time using CV at a hanging mercury drop electrode. Electrode potential-controlled adsorption driven process allowed sensitive determination of dC(AZP) at nanomolar concentrations using adsorptive stripping voltammetry. Transfer stripping voltammetry (TSV) was used for the detection of dC(AZP)-labeled DNA in femtomole quantities. Precise sequence-specific incorporation of dC(AZP) into DNA by primer extension was used to demonstrate a perfect correlation between the number of incorporated AZP moieties and TSV responses. In addition, for the first time we used polymerase chain reaction to prepare an about 350-bp double-stranded DNA fragment globally modified with dC(AZP), and of terminal deoxynucleotidyl transferase tailing reaction to generate end-labeled single stranded oligonucleotides. Effects of DNA structure on the AZP-modified DNA TSV responses are discussed. (C) 2016 Elsevier Ltd. All rights reserved.
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