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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).
DNA značená aminofenylem a nitrofenylem. Syntéza pomocí enzymatické inkorporace modifikovaných nukleosid trifosfátů, studium elektrochemických vlastností takto značené DNA
Cahová, Hana ; Havran, Luděk ; Horáková Brázdilová, Petra ; Pivoňková, Hana ; Fojta, Miroslav ; Hocek, Michal
We employed single step aqueous phase Suzuki-Miyaura cross-coupling reactions of halogenated 2'-deoxynucleoside 5'-triphosphates with 3-aminophenyl and 3-nitrophenylboronic acid for synthesis of modified nucleoside triphosphates (dNTPs). These dNTPs were then enzymatically incorporated into DNA in Primer extension experiment (PEX). Electrochemical detection using square-wave voltammetry of single-strand modified oligonucleotides (ONs) bearing 3-aminophenyl and 3-nitrophenyl tag demonstated excellent utilization in labeling of DNA.

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