|
|
Primer Extension Reaction at the Gold Electrode Surface
Vidláková, Pavlína ; Havran, Luděk ; Fojta, Miroslav
One option of covalent DNA labeling is incorporation of chemically modified nucleotides using corresponding deoxynucleotide triphosphates into DNA molecules using polymerase-catalyzed primer extension. Primer extension is usually performed in solution and then the product is analyzed, often after a suitable separation. In our study we tested the possibility to prepare oligonucleotides with enzymatically incorporated anthraquinone labeled nucleotides using primer extension on templates immobilize at a gold electrode surface.
|
|
|
Electrochemical Analysis of Chemically Modified DNA at Gold Electrodes
Vidláková, Pavlína ; Havran, Luděk ; Fojta, Miroslav
DNA labeling is used to increase sensitivity of its electrochemical detection. Covalently bound markers based on a complex of osmium tetroxide with 22-bipyridine(Os,bipy) is one of those frequently applied. Os bipy reacts with pyrimidines (mainly thymines) in single stranded regions of DNA, therefore it represents a useful marker for the analysis of DNA structure or labeling of hybridization probes . Another possibility of DNA labeling is the incorporation of chemically modified dNTPs into DNA molecules with using primer extension. DNA labeling in connection with voltammetry at gold electrodes has application potential in development of sensors for studying of DNA structure and interaction.
|
|
|
Electrochemically Switchable DNA Tags
Vidláková, Pavlína ; Havran, Luděk ; Fojta, Miroslav
DNA labelling is used for the improvement of sensitivity and selectivity of its electrochemical detection. Attachment of a new electroactive group into DNA strand can give rise to a new electrochemical signal. For analysis of label DNA is often advantageous to use redox tags giving reversible redox signal because of the possibility of performing several repeated measurements with one sample. Some redox tags give reversible redox signal directly in their introduced forms. Reduction/oxidation of other redox tags is irreversible but reduction/oxidation of these groups can give rise to new groups that yield reversible electrochemistry.
|
|
|
Novel Reactive Groups for Modification DNA by Oxoosmium Complexes
Havran, Luděk ; Vidláková, Pavlína ; Špaček, Jan ; Hermanová, Monika ; Fojta, Miroslav
DNA is naturally electroactive moleclue producing several intrinsie voltammetric signals. Some of them found appliction in electrochemical analysis of DNA interactions and damage. In particular types of applictaions is a suitable use DNA labelling by redox active tags to increase selectivity and sensitivity of analysis.One type of long-term used tags are complexes of osmium tetroxide with nittrogen ligunds (Os,L). These complexes preferentially react with pyrimidines in single stand DNA producing electractive adducts. primary reaction site for Os,L is C-C double bond in pyrimidine nucleobases. In this contribution will be presented results acquired with DNA containing chemically modified nuclebase.
|
|
|
Limitations and Possibilities of Nanostructurized Silver Amalgam
Daňhel, Aleš ; Šebest, Peter ; Vidláková, Pavlína ; Ligmajer, Filip
Variability of silver amalgam offers a row of opportunities for its application in electrochemical and spectroelectrochemical methods, thanks to its chemical, mechanical, electrochemical and spectrometric properties. This work briefly revews preparation possibilities and limitations of nanostructurized crystalas and nanopartices pf silver amalgam at different forms of carbon metals (Ag, Au, Pt) and transparent materials (ITO, FTO). These detection sysytems are in the scope of ongoing systematics research with the potential to indevelopment of novel working (micro)electrodes and spectroelectrochemical cells, potentially utilizable in modern (bio)analytical applictions.
|
|
|
Electrochemical Deposition of Silver Amalgam Particles on Pyrolitic Graphite Electrode
Šebest, Peter ; Havranová-Vidláková, Pavlína ; Ligmajer, Filip ; Fojta, Miroslav ; Daňhel, Aleš
Silver solid amalgam represents suitable alternative electrode material to metallic mercury, Controlled electrodeposition of variable silver amalgam particles (AgAP) on the surface of pyrolitic graphite electrode from an electrolyte containing Agt It and Hg(II) ions is reported as a novel method suitable lot preparation of perspective supporting materials Scanning electron microscopy including energy-disperse X-ray spectrometer and image processing software were used for analysis of composition and morphology of AgAP on POE A model electrochemically reducible 4-niirophenol was used to test a potential applicability of AgAp-PGE in electroanalytical methods.
|
|
| |
|
|
Preparation and Characterization of Silver Amalgam Particles on Gold Electrode Surface
Havranová-Vidláková, Pavlína ; Šebest, Peter ; Ligmajer, Filip ; Fojta, Miroslav ; Daňhel, Aleš
This work is focused on the preparation of silver amalgam particles by its direct electrodeposition on gold surfaces using double pulse chronoamperometry from solution containing soluble Ag+ and Hg2+ ionts. During the electrodepositions of silver amalgam particles, different: i) parameters of chronoamperometry, ii) Ag/Hg ratio in solution and iii) gold electrodes were studied. All the prepared amalgam particles were characterized by scanning electron microscope with energy dispersive X-ray spectroscopy and electrochemical methods. Voltammetric application of the gold electrodes decorated by sliver amalgam particles was confirmed by analysis of model organic nitrocompound using cyclic voltammetry.
|
|
|
Electrochemical Study of Osmium Tetroxide Comptests Reactivity to DNA Bearing Butylacrylate
Havran, Luděk ; Havranová-Vidláková, Pavlína ; Špaček, Jan ; Vítová, Lada ; Hermanová, Monika ; Fojta, Miroslav
Coniplexes of osmium tetroside with nitrogen ligands (Os,L e.g.. with 2,2'-bipyridine (bpy) find application in redox labelling of DNA.. probing of DNA structure, and in studies of DNA interaction with other molecules Os,L preferentially react with pyrimidines in single strand DNA producing electroactive adducts Primary reaction site for Os,L is C=C double bond in pyrimidine nucleobases. In this contribution we introduce a new two-step technique of DNA modification with Os,bpy, consisting in enzymatic construdion or DNA beating butyl acrylate (BA) moieties attached to uracil or T-deaza adenine, followed by chemical modification of a reactive C=C double bond in BA residue.
|
|
|
Voltametrická analýza nukleosidtrifosfátů a oligonukleotidů značených antrochinonem na zlatých elektrodách
Vidláková, Pavlína ; Balintová, Jana ; Havran, Luděk ; Hocek, Michal ; Fojta, Miroslav
Elektrochemická aktivita nukleových kyselin byla objevena v 50. letech 20. století a od té doby je používána ke studiu struktury a interakcí přirozených i modifikovaných molekul nukleových kyselin i syntetických oligonukleotidů. Přestože je přirozená DNA sama o sobě elektrochemicky aktivní a je možné ji studovat na různých typech elektrod, je pro řadu analytických aplikací praktické použít DNA značenou elektroaktivními molekulami (například komplexy přechodných kovů, amino- nebo nitroskupinami, antrachinonem apod.). Tyto látky podléhají redoxním reakcím a dávají takto modifikované DNA nové elektrochemické vlastnosti, které je možné využít pro studium struktury a interakcí nukleových kyselin a v oblasti DNA diagnostiky.
|
host ::
RSS.