Team:Aachen/Project/FRET Reporter
From 2014.igem.org
(→Determining the Quenching ability of REACh1 and REACh2) |
m (→Comparing the kinetic of the double plasmid systems K1319013 + K1319008 and K1319014 + K1319008 with standard GFP expression) |
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<a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#fluorescence" style="color:black"> | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#fluorescence" style="color:black"> | ||
- | <div class="menusmall-item menusmall-info" ><div class="menukachel" style="line-height: 1.5em | + | <div class="menusmall-item menusmall-info" ><div class="menukachel" style="top:32%; line-height:1.5em;">A Faster Answer</div></div> |
<div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/0/0b/Aachen_14-10-13_GFP_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/0/0b/Aachen_14-10-13_GFP_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | ||
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<a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#fret" style="color:black"> | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#fret" style="color:black"> | ||
- | <div class="menusmall-item menusmall-info" ><div class="menukachel" style="line-height: 1.5em | + | <div class="menusmall-item menusmall-info" ><div class="menukachel" style="top:32%; line-height:1.5em;">The FRET System</div></div> |
<div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/5/54/Aachen_14-10-13_FRET_Arrows_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/5/54/Aachen_14-10-13_FRET_Arrows_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | ||
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<li style="width: 156px;margin-left: 8px;margin-right: 8px;"> | <li style="width: 156px;margin-left: 8px;margin-right: 8px;"> | ||
<a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#darkquencher" style="color:black"> | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#darkquencher" style="color:black"> | ||
- | <div class="menusmall-item menusmall-info" ><div class="menukachel" style="line-height: 1.5em | + | <div class="menusmall-item menusmall-info" ><div class="menukachel" style="top:32%; line-height:1.5em;">REACh Quenchers</div></div> |
<div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/6/65/Aachen_14-10-13_REACh_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/6/65/Aachen_14-10-13_REACh_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | ||
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<li style="width: 156px;margin-left: 8px;margin-right: 8px;"> | <li style="width: 156px;margin-left: 8px;margin-right: 8px;"> | ||
<a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#gfp-reach" style="color:black"> | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#gfp-reach" style="color:black"> | ||
- | <div class="menusmall-item menusmall-info" ><div class="menukachel" style="line-height: 1.5em | + | <div class="menusmall-item menusmall-info" ><div class="menukachel" style="top:32%; line-height:1.5em;">The Fusion Protein</div></div> |
<div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/0/02/Aachen_14-10-13_Fusion_Protein_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/0/02/Aachen_14-10-13_Fusion_Protein_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | ||
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<a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#tevprotease" style="color:black"> | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#tevprotease" style="color:black"> | ||
- | <div class="menusmall-item menusmall-info" ><div class="menukachel" style="line-height: 1.5em | + | <div class="menusmall-item menusmall-info" ><div class="menukachel" style="top:32%; line-height:1.5em;">TEV Protease</div></div> |
<div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/d/dc/Aachen_14-10-13_TEV_Protease_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/d/dc/Aachen_14-10-13_TEV_Protease_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | ||
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<a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#reachachievements" style="color:black"> | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#reachachievements" style="color:black"> | ||
- | <div class="menusmall-item menusmall-info" ><div class="menukachel" style="line-height: 1.5em | + | <div class="menusmall-item menusmall-info" ><div class="menukachel" style="top:32%; line-height:1.5em;">Achieve-<br/>ments</div></div> |
<div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/e/ef/Aachen_14-10-15_Medal_Cellocks_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/e/ef/Aachen_14-10-15_Medal_Cellocks_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | ||
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<a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#reachoutlook" style="color:black"> | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#reachoutlook" style="color:black"> | ||
- | <div class="menusmall-item menusmall-info" ><div class="menukachel" style=" | + | <div class="menusmall-item menusmall-info" ><div class="menukachel" style="top:40%;">Outlook</div></div> |
<div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/6/67/Aachen_14-10-16_Outlook_Cellocks_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/6/67/Aachen_14-10-16_Outlook_Cellocks_iNB.png); norepeat scroll 0% 0% transparent; background-size:100%"> | ||
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{{Team:Aachen/Figure|align=center|Aachen 14-10-17 IPTG REACh iFG.png|title= Composition of our biosensor for IPTG|subtitle=To test the funtionality of our sensor concept, we expressed the TEV protease with a T7 promoter.|width=900px}} | {{Team:Aachen/Figure|align=center|Aachen 14-10-17 IPTG REACh iFG.png|title= Composition of our biosensor for IPTG|subtitle=To test the funtionality of our sensor concept, we expressed the TEV protease with a T7 promoter.|width=900px}} | ||
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{{Team:Aachen/BlockSeparator}} | {{Team:Aachen/BlockSeparator}} | ||
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To better evaluate the fluorescence, the observed Optical desity (OD) was taken into account in order to achieve a fluorescence measurement independent of the amount of cells present. This way, the measurement represents the amount of fluorescence per cell only. | To better evaluate the fluorescence, the observed Optical desity (OD) was taken into account in order to achieve a fluorescence measurement independent of the amount of cells present. This way, the measurement represents the amount of fluorescence per cell only. | ||
- | {{Team:Aachen/Figure|Aachen 16-10-14 Graph2iFG.PNG|title=Comparison of K1319013 + K1319008, K1319014 + K1319008, I20260 (positive control) and B0015 (negative control)|subtitle=Both double plasmid construct exhibit a clear fluorescence signal when induced.|width= | + | {{Team:Aachen/Figure|Aachen 16-10-14 Graph2iFG.PNG|title=Comparison of K1319013 + K1319008, K1319014 + K1319008, I20260 (positive control) and B0015 (negative control)|subtitle=Both double plasmid construct exhibit a clear fluorescence signal when induced.|width=700px}} |
The negative control B0015 did not exhibit any significant fluorescence. The positive control I20260 showed a steady level of fluorescence as expected due to the constitutive expression of GFP. As expected, the production is also independent of addition of IPTG therefore the triplicates have been merged together. | The negative control B0015 did not exhibit any significant fluorescence. The positive control I20260 showed a steady level of fluorescence as expected due to the constitutive expression of GFP. As expected, the production is also independent of addition of IPTG therefore the triplicates have been merged together. | ||
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|<html> <img src="https://static.igem.org/mediawiki/2014/1/1a/Aachen_K13%2B8%2CK14%2B8%2CK731_slower_reduced.gif" width="480px"></html> | |<html> <img src="https://static.igem.org/mediawiki/2014/1/1a/Aachen_K13%2B8%2CK14%2B8%2CK731_slower_reduced.gif" width="480px"></html> | ||
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- | |'''{{{title|K1319013 + K1319008, K1319014 + K1319008 and K731520 in an non-induced (top) and induced (bottom) chip}}}'''<br />{{{subtitle|Comparing the factor of fluorescence | + | |'''{{{title|K1319013 + K1319008, K1319014 + K1319008 and K731520 in an non-induced (top) and induced (bottom) chip}}}'''<br />{{{subtitle|Comparing the factor of fluorescence increase of induced (bottom) and not induced (top) sensor chips to the first frame of the measurement. The tested constructs are K1319013 + K1319008, K1319014 + K1319008 and K731520. After five hours, the fluorescence in our dual-plasmid REACh construct contaning K1319014 had increased by a factor of about 8, while K731520 fluorescence increased by only a factor of 2-3.}}} |
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Latest revision as of 03:47, 18 October 2014
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