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

0464800 - BFÚ 2017 RIV GB eng J - Journal Article
Daňhel, Aleš - Trošanová, Zuzana - Balintová, Jana - Havran, Luděk - Hocek, Michal - Barek, J. - Fojta, Miroslav
Voltammetric analysis of 5-(4-Azidophenyl)-2 '-deoxycytidine nucleoside and azidophenyl-labelled single- and double-stranded DNAs.
Electrochimica acta. Roč. 215, OCT2016 (2016), s. 72-83. ISSN 0013-4686. E-ISSN 1873-3859
R&D Projects: GA ČR GBP206/12/G151
Institutional support: RVO:68081707 ; RVO:61388963
Keywords : Aromatic Azide * Enzymatic Incorporation * Mercury Electrode
OECD category: Electrochemistry (dry cells, batteries, fuel cells, corrosion metals, electrolysis); Organic chemistry (UOCHB-X)
Impact factor: 4.798, year: 2016
Method of publishing: Limited access
https://www.sciencedirect.com/science/article/pii/S0013468616318205?via%3Dihub

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.
Permanent Link: http://hdl.handle.net/11104/0263575