National Repository of Grey Literature 11 records found  1 - 10next  jump to record: Search took 0.00 seconds. 
Synthesis of functionalized nucleosides and nucleotides and their incorporation into DNA
Balintová, Jana ; Hocek, Michal (advisor) ; Stiborová, Marie (referee) ; Bartošík, Martin (referee)
This PhD thesis describes the synthesis of modified DNA containing electrochemical active labels such as anthraquinone, benzofurazane, azide and nitrophenyltriazole with the application in bioanalysis. The methodology of the construction of base-modified triphosphates based on Suzuki-Miyaura coupling of halogenated deoxynucleoside triphosphates with arylboronic acids, Sonogashira coupling of dNTPs with terminal acetylenes or triphosphorylation of modified nucleosides was developed. Enzymatic incorporation of functionalized dNTPs into DNA by primer extension experiment and the electrochemical properties of DNA bearing redox labels were studied by cyclic and square- wave voltammetry. At first anthraquinone was tested as an electrochemical label. Modified 2'-deoxynucleoside triphosphates bearing anthraquinone attached through an acetylene or propargylcarbamoyl linker at 5-position of 2'-deoxycytidine 5'-triphosphate and at the 7-position of 7-deaza-2'-deoxyadenosine-5'-triphosphate were prepared by Sonogashira cross-coupling. Enzymatic incorporation of the anthraquinone-labeled dNTPs into DNA by primer extension has been developed. The electrochemical properties of the anthraquinone- labeled nucleosides, dNTPs and DNA were studied by cyclic voltammetry which shows a reversible couple of peaks around...
Searching for Electrochemical Reduction Mechanism of Azidophenyl DNA Labels
Daňhel, Aleš ; Trošanová, Zuzana ; Balintová, Jana ; Hocek, Michal ; Fojta, Miroslav
This contribution brings new information to sporadic scientific results concerned to electrochemical reduction of aromatic azides. Selected compounds used as perspective DNA labels or their structural constituents such as, 4-azidophenyltrifluoroboronic acid, 4-azidophenyl modified deoxycytidine and next probable analogous compounds and/or products of their reduction, were voltammetrically studied and compared to each other in order to reveal a mechanism of their electrochemical reduction at mercury electrodes in aqueous media. Preliminary results obtained by cyclic voltammetry at mercury and carbon based electrodes and by mass spectrometry of the products isolated from batch electrolysis on mercury pool by preparative chromatography are discussed.
Voltammetric Analysis of Anthraquinone-labeled Nucleotide Triphosphates and Oligonucleotides at Gold Electrodes
Vidláková, Pavlína ; Balintová, Jana ; Havran, Luděk ; Hocek, Michal ; Fojta, Miroslav
DNA labelling is used for increase of sensitivity of electrochemical detection. When the DNA or oligonucleotides (ONs) are chemically modified, their electrochemical responses can be changed depending on electrochemical activity of the introduced moiety. One option of DNA labeling is incorporation of chemically modified nucleotides using corresponding deoxynucleotide triphosphates (dNTPs) into DNA molecules with using primer extension (PEX). In our study we used anthraquinone labeled dNTPs and ONs with enzymatically incorporated anthraquinone labeled nucleotides. Thus synthesized ON stretch bearing the anthraquinone tags was electrochemically analyzed using voltammetry at gold electrodes.
Redox DNA Labeling - from Simple DNA Detection by Osmium Tetroxide Complexes Modification to Ratiometric Sequence Analysis
Havran, Luděk ; Balintová, Jana ; Vidláková, Pavlína ; Macíčková-Cahová, Hana ; Pivoňková, Hana ; Hocek, Michal ; Fojta, Miroslav
DNA is electroactive molecule producing analytically useful intrinsic voltammetric signals. For some applications is a useful use redox active tag to improve specificity of the analysis. One from approaches how to prepare DNA bearing redox label is its modification by complexes of osmium tetroxide with nitrogen ligands. This method find wide used in highly sensitive DNA detection and in development of electrochemical DNA hybridization sensors. Another possibility for preparation of redox labeled DNA is incorporation of redox tags modified deoxynucleotide triphosphates by DNA polymerases. In this contribution will be demonstrated application of various redox labels in electrochemical analysis of DNA.
Magnetic Beads-Based Electrochemical Techniques for DNA-Protein Interaction Monitoring
Fojta, Miroslav ; Pivoňková, Hana ; Němcová, Kateřina ; Horáková Brázdilová, Petra ; Havran, Luděk ; Orság, Petr ; Vidláková, Pavlína ; Macíčková-Cahová, Hana ; Balintová, Jana ; Hocek, Michal
Electrochemical techniques, in connection with separation of nucleoprotein complexes at magnetic beads, are suitable for the monitoring of DNA-protein interactions. For the detection of complexes captured at the beads it is possible to utilize intrinsic electrochemical activity of the protein, intrinsic structure-selective signals of the DNA, or indicator DNA substrates tail-labeled with electroactive moieties.
Using Anthraquinone Labeled Nucleotide Triphosphates for Electrochemical Analysis of DNA
Vidláková, Pavlína ; Balintová, Jana ; Horáková Brázdilová, Petra ; Havran, Luděk ; Orság, Petr ; Fojta, Miroslav ; Hocek, Michal
Base-modified anthraquinone-nucleotide triphosphates were used for DNA labeling. Anthraquinone modified nucleotides were incorporated into DNA by primer extension with KOD polymerase. In cyclic and square wave voltammetry at HMDE or PGE antraquinone gave a well developed characteristic signal suitable for differentiation of the labeled DNA from unlabeled one.
Electrochemical analysis of DNA using switchable redox moieties
Fojta, Miroslav ; Daňhel, Aleš ; Horáková Brázdilová, Petra ; Plucnara, Medard ; Pivoňková, Hana ; Havran, Luděk ; Vidláková, Pavlína ; Raindlová, Veronika ; Balintová, Jana ; Macíčková-Cahová, Hana ; Hocek, Michal
Labelling of DNA with electrochemically active moieties proved to be a convenient way to the development of electrochemical techniques for the sequence-specific DNA sensing. Through combinations of various labels differing in redox potentials, independent redox coding of different DNA sequences or individual nucleobases can be attained. Applications possibilities of electrochemistry in analysis of modified DNAs are further extended by facile monitoring of chemical conversion of reactive groups on DNA during post-labelling with ultimate redox labels. In addition, controlled in situ electrochemical conversions of specific intrinsic and extrinsic DNA components can be utilized to switch their electrochemical signals and improve signal resolution.
Redox labeling and redox coding of nucleic acids
Balintová, Jana ; Brázdová, Marie ; Havran, Luděk ; Fojta, Miroslav ; Hocek, Michal
Modified nucleoside triphosphates (dNTPs) bearing benzofurazane, azides and triazoles were prepared by Sonogashira or Suzuki cross-coupling. Polymerase incorporations of the functionalyzed dNTPs into DNA by primer extension and their electrochemical properties have been studied.
Polymerase synthesis of base-modified DNA: New methods and new applications
Balintová, Jana ; Daďová, Jitka ; Kielkowski, Pavel ; Ménová, Petra ; Vaníková, Zuzana ; Riedl, Jan ; Raindlová, Veronika ; Fojta, Miroslav ; Hocek, Michal
Diverse base-modified oligonucleotides and double-stranded DNA molecules were prepared by polymerase incorporation of modified nucleoside triphosphates. The methods include primer extension, PCR, nicking enzyme amplification reaction and mixed incorporations. The modified nucleic acids were used in redox and fluorescence labelling and coding, as well as regulation of binding and cross-linking with proteins.
Synthesis of nucleosides and nucleoside triphosphates bearing anthraquinone substituents as redox probes and their enzymatic incorporation to DNA
Balintová, Jana ; Havran, Luděk ; Fojta, Miroslav ; Hocek, Michal
Modified 2′-deoxynucleosides and nucleoside triphosphates (dNTPs) bearing anthraquinone (AQ) via acetylene or propargylcarbamoyl linkers were prepared by single-step Sonogashira cross-coupling reactions halogenated nucleosides (7-iodo-7-deaza-2′-deoxyadenosine and 5-iodo-2′-deoxycytidine) or dNTPs with N-(2-propynyl)-anthraquinone carboxamide and 2-ethynylantraquinone. Polymerase incorporation of the AQ-labelled dNTPs into DNA has also been studied. Square-wave voltammetry of the AQ-labelled nucleosides and nucleotides showed one reversible peak at –0.5 V.

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