Národní úložiště šedé literatury Nalezeno 7 záznamů.  Hledání trvalo 0.10 vteřin. 
New oxidative labels for electrochemical detection of DNA
Simonova, Anna ; Havran, Luděk ; Fojta, Miroslav ; Hocek, Michal
New dCTP and dATP derivatives bearing oxidizable labels have been synthesized and incorporated to DNA for studying their electrochemical properties in different sequences of DNA.
Preparation of modified oligonucleotides by nicking enzyme amplification reaction
Ménová, Petra ; Hocek, Michal
A method for the enzymatic synthesis of short (10-22 nt) base-modified oligonucleotides was developed, based on the nicking enzyme amplification reaction. The methodology, including product isolation and purification, was scaled up to nanomolar amounts. The modified oligonucleotides were successfully used as primers in primer extension and PCR, affording primer-modified oligonucleotides and DNA. Two simple and efficient methods for fluorescent labelling of the PCR products were also developed.
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.
Tail labelled oligonucleotide probes for the detection of DNA-protein interactions
Pivoňková, Hana ; Němcová, Kateřina ; Horáková Brázdilová, Petra ; Havran, Luděk ; Macíčková-Cahová, Hana ; Hocek, Michal ; Fojta, Miroslav
DNA-protein interactions can be monitored via different ways. We introduce novel, fast and simple approaches in DNA-protein interaction detection based on electrochemical measurements of DNA alone (structure-sensitive DNA sensing), or DNA modified with osmium tetroxide bearing nitrogenous ligands, or measurements of redox-active moieties enzymatically attached to the end of a DNA substrate thus forming a labeled tail (by terminal transferase).
Redox labelling of nucleic acids for analyzing nucleotide sequences and monitoring DNA-protein interactions
Fojta, Miroslav ; Havran, Luděk ; Horáková Brázdilová, Petra ; Pivoňková, Hana ; Kostečka, Pavel ; Macíčková-Cahová, Hana ; Raindlová, Veronika ; Vrábel, Milan ; Hocek, Michal
Nucleobase labelling of DNA for electrochemical sensing was attained through chemical modification of thymine bases with osmium tetroxide in the presence of nitrogenous ligands, or via enzymatic incorporation of nucleotide conjugates with redox-active moieties using labelled deoxynucleoside triphosphates. DNA hybridization, primer extension and PCR techniques were used for sequence-specific DNA assays. Tail-labelled DNA substrates were applied to monitor DNA binding by tumour suppressor p53 protein.
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.
Polymerase construction of base-modified DNA for chemical biology
Hocek, Michal ; Macíčková-Cahová, Hana ; Kielkowski, Pavel ; Raindlová, Veronika ; Kalachová, Lubica ; Riedl, Jan ; Balintová, Jana ; Ménová, Petra
An efficient two-step methodology for construction of base-modified DNA was developed based on aqueous-phase cross-coupling reactions of halogenated nucleoside triphosphates (dNTPs) followed by polymerase incorporation. A number of diverse chemical modifications have been successfully incorporated in this way for the use in chemical biology (modulation of cleavage by restriction enzymes, interactions with diverse DNA-binding proteins and bioconjugations).

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