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

Ales Danhel, Zuzana Trosanova, Jana Balintova, Ludek Havran, Michal Hocek, Jiri Barek, Miroslav Fojta

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Voltammetric determination of a redox labeled nucleoside 5-(4-azidophenyl)-2′-deoxycytidine (dCAZP) and various polymerase-synthesized dCAZP-labeled DNAs in aqueous buffers is presented. Influence of: i) pH (2–12), ii) scan rates (0.02–10 V s−1), and iii) dCAZP concentration (0.02–10 μmol l−1), on voltammograms of dCAZP 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 dCAZP at nanomolar concentrations using adsorptive stripping voltammetry. Transfer stripping voltammetry (TSV) was used for the detection of dCAZP-labeled DNA in femtomole quantities. Precise sequence-specific incorporation of dCAZP 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 dCAZP, 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.

Original languageEnglish
Pages (from-to)72-83
Number of pages12
JournalElectrochimica Acta
Volume215
DOIs
Publication statusPublished - 10 Oct 2016
Externally publishedYes

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Keywords

  • Aromatic Azide
  • Enzymatic Incorporation
  • Mercury Electrode
  • Nucleic Acid
  • Redox Labeling
  • Voltammetry

ASJC Scopus subject areas

  • Chemical Engineering(all)
  • Electrochemistry

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