Difference between revisions of "Template:Heidelberg/ivt/abstract"
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The specific monitoring of small molecules in biochemical reactions is problem that scientist tried to solve since many years. In this project we developed <b>s</b>mall <b>m</b>olecule <b>s</b>ensor (<b>SMS</b>) enabling us to analyze biochemical reactions in <b>real-time</b> using a fluorescent readout. Using this innovative method, we were able to analyze classical <i>in vitro</i> transcriptions in detail by monitoring the ATP consumption as well as RNA strand synthesis simultaneously. Using a Spinach RNA Aptamer fused to an ATP-binding Aptamer RNA we can specifically sense ATP concentrations in real-time. Our implemented <b>JAWS software</b> generates us the best <b>ATP-binding Aptamer</b>. Thus we can even detect small changes in the concentration of ATP during RNA synthesis. To validate the JAWS software as well as to show the general feasibility of our fluorescent tool-box system we analyzed transcription efficiencies of different RNA polymerases, the influence of the buffer as well as the effect of inhibitors like heparin on transcription. | The specific monitoring of small molecules in biochemical reactions is problem that scientist tried to solve since many years. In this project we developed <b>s</b>mall <b>m</b>olecule <b>s</b>ensor (<b>SMS</b>) enabling us to analyze biochemical reactions in <b>real-time</b> using a fluorescent readout. Using this innovative method, we were able to analyze classical <i>in vitro</i> transcriptions in detail by monitoring the ATP consumption as well as RNA strand synthesis simultaneously. Using a Spinach RNA Aptamer fused to an ATP-binding Aptamer RNA we can specifically sense ATP concentrations in real-time. Our implemented <b>JAWS software</b> generates us the best <b>ATP-binding Aptamer</b>. Thus we can even detect small changes in the concentration of ATP during RNA synthesis. To validate the JAWS software as well as to show the general feasibility of our fluorescent tool-box system we analyzed transcription efficiencies of different RNA polymerases, the influence of the buffer as well as the effect of inhibitors like heparin on transcription. | ||
| − | Finally, the combination of the JAWS Software and the fluorescent readout enables the scientific community the possibility to target specifically any small molecule of interest <i>in vivo</i> and <i>in vitro</. | + | Finally, the combination of the JAWS Software and the fluorescent readout enables the scientific community the possibility to target specifically any small molecule of interest <i>in vivo</i> and <i>in vitro</i>. |
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Latest revision as of 15:29, 18 September 2015
Abstract
The specific monitoring of small molecules in biochemical reactions is problem that scientist tried to solve since many years. In this project we developed small molecule sensor (SMS) enabling us to analyze biochemical reactions in real-time using a fluorescent readout. Using this innovative method, we were able to analyze classical in vitro transcriptions in detail by monitoring the ATP consumption as well as RNA strand synthesis simultaneously. Using a Spinach RNA Aptamer fused to an ATP-binding Aptamer RNA we can specifically sense ATP concentrations in real-time. Our implemented JAWS software generates us the best ATP-binding Aptamer. Thus we can even detect small changes in the concentration of ATP during RNA synthesis. To validate the JAWS software as well as to show the general feasibility of our fluorescent tool-box system we analyzed transcription efficiencies of different RNA polymerases, the influence of the buffer as well as the effect of inhibitors like heparin on transcription. Finally, the combination of the JAWS Software and the fluorescent readout enables the scientific community the possibility to target specifically any small molecule of interest in vivo and in vitro.