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Electrochemical analysis of RNA: development of a method suitable for the characterization of products of non-enzymatic polymerization of cyclic nucleoside monophosphates under conditions modeling pr
Hesko, Ondrej ; Vacek,, Jan (referee) ; Špaček,, Jan (advisor)
This thesis focuses on the optimazation of the electrochemical method, which characterizes products of untemplated nonenzymatic polymerization of 3',5' -cyclic guanosine monophosphate (cGMP) under conditions modeling prebiotic environment. An adsorptive transfer stripping techniques on carbon electrode and gel electrophoresis were used. The method was optimized on the model system of oligonucleotides located in solution of cGMP on carbon electrode, where DNA and RNA adsorb. This technique allows simple removing of interfering substances such as cGMP, which are not present in the original sample, but they do not adsorb on the surface of electrode or they adsorb weaker than oligonucleotides or polynucleotides. Analyses are based on the selective desorption of cGMP from the surface of the carbon electrode by the chemical and physical methods before the measurement of linear voltammetry itself. Detergents, such as SDS, Tween 20 and Triton x-100 with different concentrations and electrostatic repulsions of cGMP with different negative potentials on the carbon electrode were used for the selective desorption of cGMP. The selective desorption of cGMP was observed for all detergents and inserted negative potentials. Used methods were compared and the most effective detergent for selective desorption of cGMP was SDS. Desorption of oligonucleotides was minimalized by inserted positive potential on washed carbon electrode in 0,01% SDS in basic medium. This optimized method was used on electrochemical analysis of preliminary samples of untemplated nonenzymatic polymerization of 3',5' -cGMP and compared to the analysis of gel electrophoresis.
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Characterization of Electrochemical Behaviour and Adsorption of Methyl Viologen on Boron Doped Diamond Electrodes Depending on Their Surface Pre-treatment
Fořtová, Lucie ; Fischer, Jan (advisor) ; Vyskočil, Vlastimil (referee)
In this study we focused on the electrochemical and adsorptive behaviour of methyl viologen on electrode of boron doped diamond (BDD) possessing different termination as result of electrochemical or mechanical pretreatment. Namely was performed anodic activation applying +2.4 V in 0.5 mol L-1 sulphuric acid and cathodic pretreatment in the same solution at -2.4 V to obtain predominantly H-terminated surface. While the O- terminated surface is relatively hydrophilic with low surface conductivity, the H-terminated surface is hydrophobic with certain surface conductivity, which results typically in faster heterogenous electron transfer (HET) kinetics. Fast HET kinetics was observed on polished surfaces, that were obtained by polishing with alumina suspension. Methyl viologen (MV), a widely used herbicide, was used as a model compound because its quasireversible redox transformations result in charge changes of the molecule which includes neutral (MV0 ) or positively charged (MV+ , MV2+ ) forms. Adsorption was connected with MV0 form. The pilot experiments were performed in aqueous media using cyclic voltammetry (CV). The results indicate that adsorption of MV occurs not only on H-terminated surface, but also on the polished and O-terminated surface. Adsorption of MV0 was observed on anodic peaks on...
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Electrochemical analysis of RNA: development of a method suitable for the characterization of products of non-enzymatic polymerization of cyclic nucleoside monophosphates under conditions modeling pr
Hesko, Ondrej ; Vacek,, Jan (referee) ; Špaček,, Jan (advisor)
This thesis focuses on the optimazation of the electrochemical method, which characterizes products of untemplated nonenzymatic polymerization of 3',5' -cyclic guanosine monophosphate (cGMP) under conditions modeling prebiotic environment. An adsorptive transfer stripping techniques on carbon electrode and gel electrophoresis were used. The method was optimized on the model system of oligonucleotides located in solution of cGMP on carbon electrode, where DNA and RNA adsorb. This technique allows simple removing of interfering substances such as cGMP, which are not present in the original sample, but they do not adsorb on the surface of electrode or they adsorb weaker than oligonucleotides or polynucleotides. Analyses are based on the selective desorption of cGMP from the surface of the carbon electrode by the chemical and physical methods before the measurement of linear voltammetry itself. Detergents, such as SDS, Tween 20 and Triton x-100 with different concentrations and electrostatic repulsions of cGMP with different negative potentials on the carbon electrode were used for the selective desorption of cGMP. The selective desorption of cGMP was observed for all detergents and inserted negative potentials. Used methods were compared and the most effective detergent for selective desorption of cGMP was SDS. Desorption of oligonucleotides was minimalized by inserted positive potential on washed carbon electrode in 0,01% SDS in basic medium. This optimized method was used on electrochemical analysis of preliminary samples of untemplated nonenzymatic polymerization of 3',5' -cGMP and compared to the analysis of gel electrophoresis.
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