http://2007.igem.org/wiki/index.php?title=Imperial/Wet_Lab/Results/ID3.1&feed=atom&action=historyImperial/Wet Lab/Results/ID3.1 - Revision history2024-03-28T17:04:45ZRevision history for this page on the wikiMediaWiki 1.16.5http://2007.igem.org/wiki/index.php?title=Imperial/Wet_Lab/Results/ID3.1&diff=47713&oldid=prevCheuk Ka Tong: /* Conclusion */2007-10-27T03:30:25Z<p><span class="autocomment">Conclusion</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Conclusion==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Conclusion==</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>These are the characteristics we have extracted</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>These are the characteristics we have extracted<ins class="diffchange diffchange-inline">:</ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''AHL sensitivity'''-5nM-1000nM the construct is sensitive</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''AHL sensitivity'''-5nM-1000nM the construct is sensitive</div></td></tr>
</table>Cheuk Ka Tonghttp://2007.igem.org/wiki/index.php?title=Imperial/Wet_Lab/Results/ID3.1&diff=47707&oldid=prevCheuk Ka Tong: /* Discussion */2007-10-27T03:29:54Z<p><span class="autocomment">Discussion</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Discussion==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Discussion==</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The results from the experiment were of fluorescence against time for varying levels of AHL. Using our calibration curves we managed to convert the fluorescence into concentration of GFPmut3b produced (figure 1.1) and also into Molecules of GFPmut3b synthesised per plasmid (figure 1.2). See <del class="diffchange diffchange-inline">the </del>how the calibration curves allowed us to do this by following this link to [[Imperial/Wet Lab/Results/Res1.3/Converting_Units| converting units]].</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>The results from the experiment were of fluorescence against time for varying levels of AHL. Using our calibration curves we managed to convert the fluorescence into concentration of GFPmut3b produced (figure 1.1) and also into Molecules of GFPmut3b synthesised per plasmid (figure 1.2). See how the calibration curves allowed us to do this by following this link to [[Imperial/Wet Lab/Results/Res1.3/Converting_Units| converting units]].</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The idea behind using molecules of GFPmut3b is to try to standardise our units for the chassis and normalise our data based upon the number of DNA plasmids present in our system. This idea is similar to that for the F2620 which uses the units GFP molecules synthesized per second per cell. Trying to normalise this data with the plasmids helps to make our data independent of the DNA used in this assay, i.e. for other constructs tested and characterized in vitro.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The idea behind using molecules of GFPmut3b is to try to standardise our units for the chassis and normalise our data based upon the number of DNA plasmids present in our system. This idea is similar to that for the F2620 which uses the units GFP molecules synthesized per second per cell. Trying to normalise this data with the plasmids helps to make our data independent of the DNA used in this assay, i.e. for other constructs tested and characterized in vitro.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Figure 1.1 shows;</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Figure 1.1 shows;</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*Functional at 25<sup>o</sup>C</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*Functional at 25<sup>o</sup>C<ins class="diffchange diffchange-inline">.</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*The output of '''GFPmut3b increases with input of AHL'''</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*The output of '''GFPmut3b increases with input of AHL'''<ins class="diffchange diffchange-inline">.</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*The system is sensitive to a range of '''5-1000nM AHL'''</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*The system is sensitive to a range of '''5-1000nM AHL'''<ins class="diffchange diffchange-inline">.</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*The GFPmut3b molecules synthesis stops at '''~<del class="diffchange diffchange-inline">300minutes</del>'''. This could be due to steady state or due to no synthesis of GFPmut3b. It is known not to be steady state because the [[Imperial/Wet_Lab/Results/Res1.4| degradation experiment]] proved degradation is negligible. Interestingly <del class="diffchange diffchange-inline">this </del>time at which synthesis stops is independent of the GFPmut3b molecules produced as all the [AHL] level off at the same time. This shows that the LuxR under the control of pTet is the major source of energy consumption. The pTet is a very strong promoter and is a big consumer of the energy of the chassis. This highlights the advantages of using the construct 2 [http://partsregistry.org/Part:BBa_J37032 pLux-GFPmut3b] that does not have the energetic burden of producing LuxR under pTet. <br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*The GFPmut3b molecules synthesis stops at '''~<ins class="diffchange diffchange-inline">300 minutes</ins>'''. This could be due to steady state or due to no synthesis of GFPmut3b. It is known not to be steady state because the [[Imperial/Wet_Lab/Results/Res1.4| degradation experiment]] proved degradation is negligible. Interestingly <ins class="diffchange diffchange-inline">the </ins>time at which synthesis stops is independent of the GFPmut3b molecules produced as all the [AHL]<ins class="diffchange diffchange-inline">s </ins>level off at the same time. This shows that the LuxR under the control of pTet is the major source of energy consumption. The pTet is a very strong promoter and is a big consumer of the energy of the chassis. This highlights the advantages of using the construct 2 [http://partsregistry.org/Part:BBa_J37032 pLux-GFPmut3b] that does not have the energetic burden of producing LuxR under pTet. <br><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Figure 1.2 shows us the following:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Figure 1.2 shows us the following:</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*The shape of the '''Transfer function''' shows a linear range of response between 5nM and 100nM AHL. This defines the thresholds of response.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*The shape of the '''Transfer function''' shows a linear range of response between 5nM and 100nM AHL. This defines the thresholds of response.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*The '''lower threshold of response''' is the AHL concentration that the construct will respond</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*The '''lower threshold of response''' is the AHL concentration that the construct will respond<ins class="diffchange diffchange-inline">.</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*The '''upper threshold of response''' is the value of AHL the system is saturated and increasing AHL will not increase the rate of GFP synthesis.<br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*The '''upper threshold of response''' is the value of AHL <ins class="diffchange diffchange-inline">at which </ins>the system is saturated and increasing AHL will not increase the rate of GFP synthesis.<br><br></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Conclusion==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Conclusion==</div></td></tr>
</table>Cheuk Ka Tonghttp://2007.igem.org/wiki/index.php?title=Imperial/Wet_Lab/Results/ID3.1&diff=47207&oldid=prevMaira: /* ''In vitro'' Testing of pTet-LuxR-pLux-GFPmut3b Construct with varying concentrations of AHL */2007-10-27T02:29:54Z<p><span class="autocomment">''In vitro'' Testing of pTet-LuxR-pLux-GFPmut3b Construct with varying concentrations of AHL</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>= ''In vitro'' Testing of pTet-LuxR-pLux-GFPmut3b Construct with varying concentrations of AHL=</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>= ''In vitro'' Testing of pTet-LuxR-pLux-GFPmut3b Construct with varying concentrations of AHL=</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><br clear="all"></ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Aims==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Aims==</div></td></tr>
</table>Mairahttp://2007.igem.org/wiki/index.php?title=Imperial/Wet_Lab/Results/ID3.1&diff=47202&oldid=prevMaira at 02:29, 27 October 20072007-10-27T02:29:39Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>__NOTOC__</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>__NOTOC__</div></td></tr>
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</table>Mairahttp://2007.igem.org/wiki/index.php?title=Imperial/Wet_Lab/Results/ID3.1&diff=46749&oldid=prevJec105: /* Conclusion */2007-10-27T01:40:28Z<p><span class="autocomment">Conclusion</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''Operating Temperature Range'''-Works at 25<sup>o</sup>c</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''Operating Temperature Range'''-Works at 25<sup>o</sup>c</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''Upper Threshold''' - 1000nM AHL</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''Upper Threshold''' - 1000nM AHL</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">This means that we have met some of the initial specifications we set out. One area we will need to carry on working on is trying to create a visual threshold using a stronger fluorescent protein.</ins></div></td></tr>
</table>Jec105http://2007.igem.org/wiki/index.php?title=Imperial/Wet_Lab/Results/ID3.1&diff=46741&oldid=prevJec105: /* Conclusion */2007-10-27T01:39:22Z<p><span class="autocomment">Conclusion</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''Life of Synthesis'''-300 minutes</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''Life of Synthesis'''-300 minutes</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''Operating Temperature Range'''-Works at 25<sup>o</sup>c</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''Operating Temperature Range'''-Works at 25<sup>o</sup>c</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*'''Upper Threshold''' - 1000nM AHL</ins></div></td></tr>
</table>Jec105http://2007.igem.org/wiki/index.php?title=Imperial/Wet_Lab/Results/ID3.1&diff=46733&oldid=prevJec105 at 01:38, 27 October 20072007-10-27T01:38:45Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>These are the characteristics we have extracted</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>These are the characteristics we have extracted</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Switch Point= ~20nM </del>AHL<del class="diffchange diffchange-inline"><br></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">*'''</ins>AHL <ins class="diffchange diffchange-inline">sensitivity'''-5nM-1000nM the construct is sensitive</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Response Time= <30 </del>minutes<<del class="diffchange diffchange-inline">br</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">*'''Life of Synthesis'''-300 </ins>minutes</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Sensitivity = </del><<del class="diffchange diffchange-inline">5nM AHL<BR</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">*'''Operating Temperature Range'''-Works at 25</ins><<ins class="diffchange diffchange-inline">sup</ins>><ins class="diffchange diffchange-inline">o</ins><<ins class="diffchange diffchange-inline">/sup</ins>><ins class="diffchange diffchange-inline">c</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">In terms of the specification of the Infector Detector Application the chassis and construct is sensitive enough.</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
</table>Jec105http://2007.igem.org/wiki/index.php?title=Imperial/Wet_Lab/Results/ID3.1&diff=46716&oldid=prevJec105: /* Discussion */2007-10-27T01:36:37Z<p><span class="autocomment">Discussion</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The idea behind using molecules of GFPmut3b is to try to standardise our units for the chassis and normalise our data based upon the number of DNA plasmids present in our system. This idea is similar to that for the F2620 which uses the units GFP molecules synthesized per second per cell. Trying to normalise this data with the plasmids helps to make our data independent of the DNA used in this assay, i.e. for other constructs tested and characterized in vitro.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The idea behind using molecules of GFPmut3b is to try to standardise our units for the chassis and normalise our data based upon the number of DNA plasmids present in our system. This idea is similar to that for the F2620 which uses the units GFP molecules synthesized per second per cell. Trying to normalise this data with the plasmids helps to make our data independent of the DNA used in this assay, i.e. for other constructs tested and characterized in vitro.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">There are several key parameters that can be extracted from the data</del>. <del class="diffchange diffchange-inline">It looks as if our system becomes saturated to AHL ( the </del>input<del class="diffchange diffchange-inline">) above 100nM </del>of AHL<del class="diffchange diffchange-inline">, it can be seen that there </del>is <del class="diffchange diffchange-inline">little difference between </del>1000nM <del class="diffchange diffchange-inline">and 100nM</del>. <del class="diffchange diffchange-inline">If we assume the GFP </del>synthesis <del class="diffchange diffchange-inline">1000nM </del>to be the <del class="diffchange diffchange-inline">maximum then the switch point , defined when the output </del>is <del class="diffchange diffchange-inline">50% </del>of the <del class="diffchange diffchange-inline">max, </del>is <del class="diffchange diffchange-inline">around 20nM </del>of <del class="diffchange diffchange-inline">AHL</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Figure 1</ins>.<ins class="diffchange diffchange-inline">1 shows;</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">*Functional at 25<sup>o</sup>C</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The <del class="diffchange diffchange-inline">response time </del>is a <del class="diffchange diffchange-inline">key characteristic however from this data it </del>is <del class="diffchange diffchange-inline">difficult to get an accurate value</del>. <del class="diffchange diffchange-inline">The problem is </del>that <del class="diffchange diffchange-inline">because </del>the <del class="diffchange diffchange-inline">temperature was regulated sampling had to be restricted to every 30 minutes, meaning our best approximation is <30minutes</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">*The output of '''GFPmut3b increases with </ins>input of AHL<ins class="diffchange diffchange-inline">'''</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">*The system </ins>is <ins class="diffchange diffchange-inline">sensitive to a range of '''5-</ins>1000nM <ins class="diffchange diffchange-inline">AHL'''</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The <del class="diffchange diffchange-inline">sensitivity </del>of the <del class="diffchange diffchange-inline">chassis </del>and <del class="diffchange diffchange-inline">construct can also be deduced for </del>the <del class="diffchange diffchange-inline">range </del>of AHL <del class="diffchange diffchange-inline">concentrations </del>that <del class="diffchange diffchange-inline">we looked at. </del>The <del class="diffchange diffchange-inline">sensitivity to </del>AHL is <del class="diffchange diffchange-inline"><5nM</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">*The GFPmut3b molecules synthesis stops at '''~300minutes'''</ins>. <ins class="diffchange diffchange-inline">This could be due to steady state or due to no </ins>synthesis <ins class="diffchange diffchange-inline">of GFPmut3b. It is known not </ins>to be <ins class="diffchange diffchange-inline">steady state because </ins>the <ins class="diffchange diffchange-inline">[[Imperial/Wet_Lab/Results/Res1.4| degradation experiment]] proved degradation is negligible. Interestingly this time at which synthesis stops </ins>is <ins class="diffchange diffchange-inline">independent </ins>of the <ins class="diffchange diffchange-inline">GFPmut3b molecules produced as all the [AHL] level off at the same time. This shows that the LuxR under the control of pTet </ins>is <ins class="diffchange diffchange-inline">the major source </ins>of <ins class="diffchange diffchange-inline">energy consumption</ins>. The <ins class="diffchange diffchange-inline">pTet </ins>is a <ins class="diffchange diffchange-inline">very strong promoter and </ins>is <ins class="diffchange diffchange-inline">a big consumer of the energy of the chassis</ins>. <ins class="diffchange diffchange-inline">This highlights the advantages of using the construct 2 [http://partsregistry.org/Part:BBa_J37032 pLux-GFPmut3b] </ins>that <ins class="diffchange diffchange-inline">does not have </ins>the <ins class="diffchange diffchange-inline">energetic burden of producing LuxR under pTet</ins>. <ins class="diffchange diffchange-inline"><br><br></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Figure 1.2 shows us the following:</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">*</ins>The <ins class="diffchange diffchange-inline">shape </ins>of the <ins class="diffchange diffchange-inline">'''Transfer function''' shows a linear range of response between 5nM </ins>and <ins class="diffchange diffchange-inline">100nM AHL. This defines </ins>the <ins class="diffchange diffchange-inline">thresholds </ins>of <ins class="diffchange diffchange-inline">response.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">*The '''lower threshold of response''' is the </ins>AHL <ins class="diffchange diffchange-inline">concentration </ins>that <ins class="diffchange diffchange-inline">the construct will respond</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">*</ins>The <ins class="diffchange diffchange-inline">'''upper threshold of response''' is the value of </ins>AHL <ins class="diffchange diffchange-inline">the system </ins>is <ins class="diffchange diffchange-inline">saturated and increasing AHL will not increase the rate of GFP synthesis</ins>.<ins class="diffchange diffchange-inline"><br><br></ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Conclusion==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Conclusion==</div></td></tr>
</table>Jec105http://2007.igem.org/wiki/index.php?title=Imperial/Wet_Lab/Results/ID3.1&diff=46676&oldid=prevJec105: /* Discussion */2007-10-27T01:31:09Z<p><span class="autocomment">Discussion</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Discussion==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Discussion==</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The results from the experiment were of fluorescence against time for varying levels of AHL. Using our calibration curves we managed to convert the fluorescence into concentration of GFPmut3b produced (figure 1.1) and also into Molecules of GFPmut3b synthesised per plasmid (figure 1.2). See the how the calibration curves allowed us to do this by following this link to <del class="diffchange diffchange-inline">conversion of </del>units <del class="diffchange diffchange-inline">page (LINK)</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>The results from the experiment were of fluorescence against time for varying levels of AHL. Using our calibration curves we managed to convert the fluorescence into concentration of GFPmut3b produced (figure 1.1) and also into Molecules of GFPmut3b synthesised per plasmid (figure 1.2). See the how the calibration curves allowed us to do this by following this link to <ins class="diffchange diffchange-inline"> [[Imperial/Wet Lab/Results/Res1.3/Converting_Units| converting </ins>units<ins class="diffchange diffchange-inline">]]</ins>.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The idea behind using molecules of GFPmut3b is to try to standardise our units for the chassis and normalise our data based upon the number of DNA plasmids present in our system. This idea is similar to that for the F2620 which uses the units GFP molecules synthesized per second per cell. Trying to normalise this data with the plasmids helps to make our data independent of the DNA used in this assay, i.e. for other constructs tested and characterized in vitro.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The idea behind using molecules of GFPmut3b is to try to standardise our units for the chassis and normalise our data based upon the number of DNA plasmids present in our system. This idea is similar to that for the F2620 which uses the units GFP molecules synthesized per second per cell. Trying to normalise this data with the plasmids helps to make our data independent of the DNA used in this assay, i.e. for other constructs tested and characterized in vitro.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The sensitivity of the chassis and construct can also be deduced for the range of AHL concentrations that we looked at. The sensitivity to AHL is <5nM.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The sensitivity of the chassis and construct can also be deduced for the range of AHL concentrations that we looked at. The sensitivity to AHL is <5nM.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"> </td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Conclusion==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Conclusion==</div></td></tr>
</table>Jec105http://2007.igem.org/wiki/index.php?title=Imperial/Wet_Lab/Results/ID3.1&diff=46664&oldid=prevJec105: /* Results */2007-10-27T01:29:36Z<p><span class="autocomment">Results</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*Sampling on same plate and every 30 minutes</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*Sampling on same plate and every 30 minutes</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>'''Raw Data'''</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>'''Raw Data'''<ins class="diffchange diffchange-inline"><br></ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>[[Image:AHL testing Final1.xls|Raw Data]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>[[Image:AHL testing Final1.xls| Raw Data]]</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Discussion==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Discussion==</div></td></tr>
</table>Jec105