Difference between revisions of "Team:CU Boulder/project/results"
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<h1>Results</h1> | <h1>Results</h1> | ||
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+ | <!--------------------Switch and GFP--------------------------> | ||
+ | <h2>Characterization of Switch with LuxI-GFP</h2> | ||
+ | <p>Preliminary results with RFP and GFP showed that the switch was leaky when on a pSB1C3 backbone. We believe that because of the high copy number of pSB1C3, there was a high number of switches in each cell so if only a small proportion of switches were to leak, this effect would be visible. In order to investigate this issue, we transfered the gate with a GFP reporter onto a low copy plasmid. As expected, we saw much less expression of our reporter.</p> | ||
+ | <br> | ||
+ | <p>To test the responsiveness of our integrase in tandem with the leakiness of the gate on a low copy plasmid we co-transformed the Bxb1 integrase behind the arabinose inducible promoter on a kanamycinbackbone with BBa_K1718005 (switch-LuxI-GFP) on 4C5.</p> | ||
+ | <p>It was tested by growing cells in overnight cultures with varying concentrations of arabinose, from 0% to .2%increasing by .04% for a total of 6 cultures. The cultures were made of 5mL of LB, 5ul of both chloramphenicol and kanamycin, and cells taken from a glycerol stock of cells with the desired plasmids. The 4th culture had low growth.</p> | ||
+ | <p>Flow cytometry was performed on the cells after being resuspended in PBS which returned the resulting data.</p> | ||
+ | <div class="frame" id="4c5_flow"></div> | ||
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+ | <p>With the exception of the fourth sample, which was the culture with low growth, we see a general rightward trend in the peaks, indicating that higher concentrations of arabinose are more effective at inducing the production of the integrase. More tests should be done to confirm this trend and find the ideal concentration.</p> | ||
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<!---------------------pBAD promoter --------------------------> | <!---------------------pBAD promoter --------------------------> |
Revision as of 08:56, 18 September 2015
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