IGEM:Imperial/2010/Experiments1: Difference between revisions
Anita Nguyen (talk | contribs) (New page: =Experiments for the Output Amplification Model= * Determine maximum concentration of sD that cells can produce – crucial! (''in vivo''): Compare activities with the next one * Determine...) |
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=Experiments for the Output Amplification Model= | <html> | ||
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Compare | <h1>Experiments for the Output Amplification Model</h1> | ||
In vivo IPTG | <img src="http://upload.wikimedia.org/wikipedia/commons/thumb/4/4f/Postit_large.jpg/500px-Postit_large.jpg"><div style="position:relative;top:-450px;left:50px;font: 11pt helvetica, georgia, courier, arial; margin-left: 10px;"><b>Output Amplification Model</b><br /><br /> | ||
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<li>Determine the maximum concentration of sD that <br />cells can produce (in vivo): | |||
TEV | Compare activity with 2. | ||
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<li>Determine how much XylE we need to produce <br />prior to adding Catechol (in vitro first, maybe in vivo <br />afterwards). In vivo IPTG: Compare cultures that <br />have less of it. | |||
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<li>Determine XylE and TEV production (in vivo). <br />Use robot to induce production and measure <br />activities TEV (FRET pairs with TEV link). | |||
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<li>Degradation of XylE (in vivo or in vitro if cell division <br />is not taken into account). Monitoring activity, then <br />approximate the concentration. Remove the IPTG. | |||
</li> | |||
<li>Determine TEV kinetics (k<sub>cat</sub>, K<sub>m</sub>) on <br />XylE-GFP (in vitro).</li> | |||
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Latest revision as of 07:27, 9 September 2010
<html> <body style="background-color:FFFFCC"> <h1>Experiments for the Output Amplification Model</h1>
<img src="http://upload.wikimedia.org/wikipedia/commons/thumb/4/4f/Postit_large.jpg/500px-Postit_large.jpg"><div style="position:relative;top:-450px;left:50px;font: 11pt helvetica, georgia, courier, arial; margin-left: 10px;"><b>Output Amplification Model</b><br /><br /> <ol> <li>Determine the maximum concentration of sD that <br />cells can produce (in vivo): Compare activity with 2. </li> <li>Determine how much XylE we need to produce <br />prior to adding Catechol (in vitro first, maybe in vivo <br />afterwards). In vivo IPTG: Compare cultures that <br />have less of it. </li> <li>Determine XylE and TEV production (in vivo). <br />Use robot to induce production and measure <br />activities TEV (FRET pairs with TEV link). </li> <li>Degradation of XylE (in vivo or in vitro if cell division <br />is not taken into account). Monitoring activity, then <br />approximate the concentration. Remove the IPTG. </li> <li>Determine TEV kinetics (k<sub>cat</sub>, K<sub>m</sub>) on <br />XylE-GFP (in vitro).</li> </ol> </div> </body> </html>
