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 language | English |
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Pages (from-to) | 72-83 |
Number of pages | 12 |
Journal | Electrochimica Acta |
Volume | 215 |
DOIs | |
Publication status | Published - 10 Oct 2016 |
Externally published | Yes |
Keywords
- Aromatic Azide
- Enzymatic Incorporation
- Mercury Electrode
- Nucleic Acid
- Redox Labeling
- Voltammetry
ASJC Scopus subject areas
- Chemical Engineering(all)
- Electrochemistry