Chronopotentiometric stripping of DNA at mercury electrodes

Paleček, Emil; Tomschik, Miroslav; Staňková, Veronika; Havran, Luděk

doi:10.1002/elan.1140091305

Cited by 51 publications

(27 citation statements)

References 37 publications

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“…[27][28][29][30][31][32] However, a cathode peak of the reduction of guanine, which was believed to be at about -1.85 V, was not observed. 33,34 The maximum cathode peak current of ds-DNA in the presence of Al(III) was 6.6% higher than that of the thermal denatured sd-DNA ( Fig.…”

Section: Electroanalytical Studies On the Denaturation Of Ds-dna By Amentioning

confidence: 95%

Electrochemical and Spectrometric Studies of Double-Strand Calf Thymus Gland DNA Denatured by Al(III) at Neutral pH

et al. 2009

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The interaction between double-strand calf thymus gland DNA (ds-DNA) and Al(III) was studied by using differential pulse voltammetry (DPV) at a hanging mercury drop electrode (HMDE), Raman spectrometry and circular dichroism (CD) spectra. It was shown that at neutral pH ds-DNA did not produce any cathodic peak at the HMDE in the potential window from -550 to -2000 mV vs. SCE. However, in the presence of Al(III), a cathodic peak was generated at about -1660 mV, which is ascribed to a reduction of adenine and cytosine residues of single denatured DNA (sd-DNA). It was concluded that ds-DNA was completely denatured to sd-DNA by Al(III) at a neutral pH. The apparent denaturing kinetic velocity constants of ds-DNA by Al(III) were derived from linear increases of the cathodic peak currents with time.3 ≥ 2 × 10 -26 , the precipitation of Al(OH)3 was observed and identified by the Raman spectrum, and inductively coupled plasma atomic emission spectrometry (ICP-AES). CD spectra showed that the B-type of structure conformations of ds-DNA and related sd-DNA did not change with the increment of Al(III) from 5.0 × 10 -7 to 1.0 10 -5 M, but the corresponding absorption strengths increased. The related physiological significances and possible applications of the observations were considered.

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Section: Electroanalytical Studies On the Denaturation Of Ds-dna By Amentioning

confidence: 95%

Electrochemical and Spectrometric Studies of Double-Strand Calf Thymus Gland DNA Denatured by Al(III) at Neutral pH

et al. 2009

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“…This technique can also be applied for the analysis of DNA and proteins (Palecek et al, 1997a;Tomschik et al, 1998). Measurements were performed with an AUTOLAB (EcoChemie, Utrecht, The Netherlands) connected to a Metrohm 647 VA Stand (mercury electrode) in HMDE mode (drop surface area 0.4 mm 2 ) at a constant current of 71 mA using 0.2 M sodium phosphate as a background electrolyte.…”

Section: Blotting With Immunodetection Of P53mentioning

confidence: 99%

Effect of transition metals on binding of p53 protein to supercoiled DNA and to consensus sequence in DNA fragments

et al. 1999

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Recently we have shown that wild-type human p53 protein binds preferentially to supercoiled (sc) DNA in vitro in both the presence and absence of the p53 consensus sequence (p53CON). This binding produces a ladder of retarded bands on an agarose gel. Using immunoblotting with the antibody DO-1, we show that the bands obtained correspond to ethidium-stained DNA, suggesting that each band of the ladder contains a DNAp53 complex. The intensity and the number of these bands are decreased by physiological concentrations of zinc ions. At higher zinc concentrations, binding of p53 to scDNA is completely inhibited. The binding of additional zinc ions to p53 appears much weaker than the binding of the intrinsic zinc ion in the DNA binding site of the core domain. In contrast to previously published data suggesting that 100 mM zinc ions do not in¯uence p53 binding to p53CON in a DNA oligonucleotide, we show that 5 ± 20 mM zinc e ciently inhibits binding of p53 to p53CON in DNA fragments. We also show that relatively low concentrations of dithiothreitol but not of 2-mercaptoethanol decrease the concentration of free zinc ions, thereby preventing their inhibitory e ect on binding of p53 to DNA. Nickel and cobalt ions inhibit binding of p53 to scDNA and to its consensus sequence in linear DNA fragments less e ciently than zinc; cobalt ions are least e cient, requiring 4100 mM Co 2+ for full inhibition of p53 binding. Modulation of binding of p53 to DNA by physiological concentrations of zinc might represent a novel pathway that regulates p53 activity in vivo.

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“…Using AdTSV involving extraction of free Os, bipy from the electrode surface by organic solvents [22], it is thus possible to distinguish between free Os, bipy and Os, bipy-labeled DNA, and to determine DNA-Os, bipy in an excess of the unreacted complex. We have shown that, with random-sequence calf thymus DNA, this technique allowed to detect about 150 picograms of DNA in several microliters of solution with neither stirring nor application of positive potentials (about the same sensitivity was reached in measurements of guanine oxidation signal at carbon electrodes, but only when the solution was stirred and DNA adsorbed at positively charged electrode surface to facilitate DNA accumulation at the electrode [33]). …”

Section: Dna Hybridization At Magnetic Beads and Dna Osmium Labelingmentioning

confidence: 99%

Two‐Surface Strategy in Electrochemical DNA Hybridization Assays: Detection of Osmium‐Labeled Target DNA at Carbon Electrodes

et al. 2003

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Target DNAs, including a 71-mer oligonucleotide, a PCR product and a plasmid DNA, all containing oligo(A) stretches, were hybridized at magnetic Dynabeads oligo(dT) 25 (DBT). The hybridization events were detected using a technique based on chemical modification of the target DNA with a complex of osmium tetroxide with 2,2'-bipyridine (Os, bipy) and voltammetric detection at carbon electrodes. DNA was modified with Os, bipy prior to capture at DBT, at the beads, or after release from the beads. In the latter case, DNA-Os, bipy was detected in the reaction mixture using adsorptive transfer stripping voltammetry involving extraction of unreacted Os, bipy from the electrode by organic solvents. Pre-labeling of the target plasmid DNA and the PCR product with Os, bipy significantly increased the yield of DNA captured at the beads. Tens of femtomoles of both short (the 71-mer oligonucleotide) and long (the 3-kilobase plasmid) target DNAs in a 20-microliter hybridization sample can be easily detected by means of these techniques. Various carbon electrode materials, including pyrolytic graphite (PGE), highly oriented pyrolytic graphite (HOPGE), carbon paste (CPE), glassy carbon and pencil graphite, were tested regarding their suitability for the detection of osmium-labeled DNA. Among them, PGE and HOPGE appeared usable in the measurements of both purified DNA-Os, bipy and its mixtures with unreacted Os, bipy while CPE was suitable for the detection purified osmium-labeled DNA.

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Chronopotentiometric stripping of DNA at mercury electrodes

Cited by 51 publications

References 37 publications

Electrochemical and Spectrometric Studies of Double-Strand Calf Thymus Gland DNA Denatured by Al(III) at Neutral pH

Electrochemical and Spectrometric Studies of Double-Strand Calf Thymus Gland DNA Denatured by Al(III) at Neutral pH

Effect of transition metals on binding of p53 protein to supercoiled DNA and to consensus sequence in DNA fragments

Two‐Surface Strategy in Electrochemical DNA Hybridization Assays: Detection of Osmium‐Labeled Target DNA at Carbon Electrodes

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