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

- Voltammetric behaviour of 5-(4-azidophenyl)-2′-deoxycytidine (dCAZP) at HMDE in aqueous media.
- Nanomolar concentrations of dCAZP and ss and dsDNA labeled by azidophenyl detectable at HMDE.
- Precise sequence-specific incorporation of dCAZP into DNA by primer extension.
- Polymerase chain reaction produces an about 350-bp double-stranded DNA fragment globally modified with dCAZP.
- Terminal deoxynucleotidyl transferase tailing reaction generates dCAZP end-labeled single stranded oligonucleotides

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.

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