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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.
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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.
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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.
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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.
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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.
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