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 |
|---|---|
| Pages (from-to) | 72-83 |
| Number of pages | 12 |
| Journal | Electrochimica Acta |
| Volume | 215 |
| DOIs | |
| Publication status | Published - 10 Oct 2016 |
| Externally published | Yes |
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Keywords
- Aromatic Azide
- Enzymatic Incorporation
- Mercury Electrode
- Nucleic Acid
- Redox Labeling
- Voltammetry
ASJC Scopus subject areas
- Chemical Engineering(all)
- Electrochemistry
Cite this
Voltammetric analysis of 5-(4-Azidophenyl)-2′-deoxycytidine nucleoside and azidophenyl-labelled single- and double-stranded DNAs. / Danhel, Ales; Trosanova, Zuzana; Balintova, Jana; Havran, Ludek; Hocek, Michal; Barek, Jiri; Fojta, Miroslav.
In: Electrochimica Acta, Vol. 215, 10.10.2016, p. 72-83.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Voltammetric analysis of 5-(4-Azidophenyl)-2′-deoxycytidine nucleoside and azidophenyl-labelled single- and double-stranded DNAs
AU - Danhel, Ales
AU - Trosanova, Zuzana
AU - Balintova, Jana
AU - Havran, Ludek
AU - Hocek, Michal
AU - Barek, Jiri
AU - Fojta, Miroslav
PY - 2016/10/10
Y1 - 2016/10/10
N2 - 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.
AB - 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.
KW - Aromatic Azide
KW - Enzymatic Incorporation
KW - Mercury Electrode
KW - Nucleic Acid
KW - Redox Labeling
KW - Voltammetry
UR - http://www.scopus.com/inward/record.url?scp=84983284263&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84983284263&partnerID=8YFLogxK
U2 - 10.1016/j.electacta.2016.08.096
DO - 10.1016/j.electacta.2016.08.096
M3 - Article
AN - SCOPUS:84983284263
VL - 215
SP - 72
EP - 83
JO - Electrochimica Acta
JF - Electrochimica Acta
SN - 0013-4686
ER -