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Team:Paris Bettencourt/Experiment/T7 diff subt subt microfluidic
From 2011.igem.org
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<h2>Experimental Scheme</h2> | <h2>Experimental Scheme</h2> | ||
| - | <p>In order to test whether our <i>Bacillus subtilis</i> T7 emitters & receivers can form nanotubes when mixed, we mix them in microfluidic system modified from Jeff Hasty's recent <a href="http://biodynamics.ucsd.edu/pubs/articles/Mondragon11.pdf">paper</a> <a href="https://2011.igem.org/Team:Paris_Bettencourt/Experiments/Methodologies/Microchemostat_HastyJ#references">[1]</a>, shown below. | + | <p>In order to test whether our <i>Bacillus subtilis</i> T7 emitters & receivers can form nanotubes when mixed, we mix them in microfluidic system modified from Jeff Hasty's recent <a href="http://biodynamics.ucsd.edu/pubs/articles/Mondragon11.pdf">paper</a> <a href="https://2011.igem.org/Team:Paris_Bettencourt/Experiments/Methodologies/Microchemostat_HastyJ#references">[1]</a>, shown below. For more detailed information regarding the microfluidic device and experimental procedure, check our <a href="https://2011.igem.org/Team:Paris_Bettencourt/Experiments/Methodologies/Microchemostat_HastyJ">methodology page</a>. </p> |
| - | <center><a href="https://2011.igem.org/File:Paris_microchemostat_channels_and_chambers.jpg"><img height=540px align="center" src="https://static.igem.org/mediawiki/2011/f/f9/Paris_microchemostat_channels_and_chambers.jpg"></a></center> | + | <p><center><a href="https://2011.igem.org/File:Paris_microchemostat_channels_and_chambers.jpg"><img height=540px align="center" src="https://static.igem.org/mediawiki/2011/f/f9/Paris_microchemostat_channels_and_chambers.jpg"></a></center></p> |
| - | We imaged two channels: one experimental channel injected with an emitter strain (RFP constitutive) and a receiver strain (pT7-T7polyermase-GFP); the other is a control channel injected with only the receiver strain. The receiver strain contains the T7 autoloop, which will gain a strong fluorescence when activated. </p> | + | <p>We imaged two channels: one experimental channel injected with an emitter strain (RFP constitutive) and a receiver strain (pT7-T7polyermase-GFP); the other is a control channel injected with only the receiver strain. The receiver strain contains the T7 autoloop, which will gain a strong fluorescence when activated. </p> |
<h2>Results</h2> | <h2>Results</h2> | ||
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| + | The receiver strain by itself in our control channel have never produced any GFP positive cells, in contrast to the leaky situation where the T7 autoloop is encoded by a plasmid in E.coli. This is due to the low copy number of T7 polymerase gene on the chromosome, | ||
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| + | <p><center><a href="https://static.igem.org/mediawiki/2011/6/69/Paris2011_T7-microfluidics-gfpsums.jpg"><img height=540px align="center" src="https://static.igem.org/mediawiki/2011/6/69/Paris2011_T7-microfluidics-gfpsums.jpg"></a></center></p> | ||
<div id="citation_box"> | <div id="citation_box"> | ||
Revision as of 03:08, 29 October 2011
Nanotube-assisted diffusion of T7 polymerase in Microfluidics
Experimental Scheme
In order to test whether our Bacillus subtilis T7 emitters & receivers can form nanotubes when mixed, we mix them in microfluidic system modified from Jeff Hasty's recent paper [1], shown below. For more detailed information regarding the microfluidic device and experimental procedure, check our methodology page.
We imaged two channels: one experimental channel injected with an emitter strain (RFP constitutive) and a receiver strain (pT7-T7polyermase-GFP); the other is a control channel injected with only the receiver strain. The receiver strain contains the T7 autoloop, which will gain a strong fluorescence when activated.
Results
The receiver strain by itself in our control channel have never produced any GFP positive cells, in contrast to the leaky situation where the T7 autoloop is encoded by a plasmid in E.coli. This is due to the low copy number of T7 polymerase gene on the chromosome, d
References
- Entrainment of a population of synthetic genetic oscillators. Mondragón-Palomino, O., Danino, T., Selimkhanov, J., Tsimring, L. & Hasty, J. Science 333, 1315-1319 (2011).
