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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 (oponent) ; Špaček,, Jan (vedoucí práce)
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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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 (oponent) ; Špaček,, Jan (vedoucí práce)
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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Evolution of life - the RNA world
Čvirik, Rastislav ; Markoš, Anton (vedoucí práce) ; Kalous, Martin (oponent)
Vznik a evolúcia života v prírodných vedách predstavuje komplexnú a doposiaľ nevyriešenú otázku. Jedno z možných vysvetlení ponúka hypotéza "sveta RNA". Jej hlavnou myšlienkou je, že molekula RNA môže fungovať ako nositeľ genetickej informácie a zároveň aj ako katalyzátor chemických reakcií. Rovnako predpokladá, že v skorej fáze vývoja života na Zemi bola genetická kontinuita zaistená replikáciou molekuly RNA, proteíny a DNA sa vynorili až neskôr. Z recentných výskumov sa vynárajú stále silnejšie dôkazy ktoré podporujú existenciu tohoto sveta. Zostáva však stále množstvo problémov a nevyriešených paradoxov spojených prebiotickou chémiou. Aj keď táto teória priamo nevysvetľuje vznik života, ponúka zaujímavú alternatívu ako k tejto udalosti mohlo dojsť na primitívnej Zemi. Powered by TCPDF (www.tcpdf.org)
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