Difference between revisions of "Team:NTU-Singapore/Results"
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<p class="subtitle explain">We did another round of measurements with additional mutants. However, for the second batch, our lactate dehydrogenase mutants were now ligated to the wildtype version of lactate permease. Other than that, we used the BioScreen which allows an automated measurements of our cultures in a 96-well plate. However, some of the mutants did not grew after inoculations. For lactate permease mutants, we did not carry on further as multiple ligations lead to failure. | <p class="subtitle explain">We did another round of measurements with additional mutants. However, for the second batch, our lactate dehydrogenase mutants were now ligated to the wildtype version of lactate permease. Other than that, we used the BioScreen which allows an automated measurements of our cultures in a 96-well plate. However, some of the mutants did not grew after inoculations. For lactate permease mutants, we did not carry on further as multiple ligations lead to failure. | ||
<div align="centre"><img class="" src="https://static.igem.org/mediawiki/2015/4/43/Chart_M9_Set1.png" width="600px" height="450px"></div><p style="text-align:center; font-size: 19px; color: black"> | <div align="centre"><img class="" src="https://static.igem.org/mediawiki/2015/4/43/Chart_M9_Set1.png" width="600px" height="450px"></div><p style="text-align:center; font-size: 19px; color: black"> | ||
| − | Growth of Shewanella oneidensis MR1 over expressing the mutant lactate dehydrogenase and the wildtype lactate permease. 21: Wild type control, 22: empty | + | Growth of Shewanella oneidensis MR1 over expressing the mutant lactate dehydrogenase and the wildtype lactate permease. 21: Wild type control, 22: empty vector. |
</p> | </p> | ||
<div align="centre"><img class="" src="https://static.igem.org/mediawiki/2015/e/e7/Chart_M9_Set2.png" width="600px" height="450px"></div><p style="text-align:center; font-size: 19px; color: black"> | <div align="centre"><img class="" src="https://static.igem.org/mediawiki/2015/e/e7/Chart_M9_Set2.png" width="600px" height="450px"></div><p style="text-align:center; font-size: 19px; color: black"> | ||
| − | Growth of Shewanella oneidensis MR1 over expressing the mutant lactate dehydrogenase and the wildtype lactate permease. 21: Wild type control, 22: empty | + | Growth of Shewanella oneidensis MR1 over expressing the mutant lactate dehydrogenase and the wildtype lactate permease. 21: Wild type control, 22: empty vector. |
</p> | </p> | ||
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<div class="inner" > | <div class="inner" > | ||
| − | <h5> | + | <h5>Microbial Fuel Cells</h5> |
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| + | <p class="subtitle explain"> When the mutants were tested for the voltage output, several mutants showed high voltage output. The measurements was made on the closed circuit potential with two resistors connecting the anode to the cathode. | ||
| + | <div align="centre"><img class="" src="https://static.igem.org/mediawiki/2015/4/43/Chart_M9_Set1.png" width="600px" height="450px"></div><p style="text-align:center; font-size: 19px; color: black"> | ||
| + | Growth of Shewanella oneidensis MR1 over expressing the mutant lactate dehydrogenase and the wildtype lactate permease. 21: Wild type control, 22: empty vector. | ||
| + | </p> | ||
</div> | </div> | ||
Revision as of 02:30, 19 September 2015
Our Results
Ribosomal Binding Site
Growth Curve
As the measurements are carried out in six batches, the growth of mutants of the same batch are similar but differed a little among batches. This implies that GFP expression does not have any significant effect on the bacteris's growth.
1AT denotes A of base pair 1 is changed to T, the same is applied for other notations of RBS mutants.
GFP Readings
The GFP fluorescence readings of mutants shows interesting results. Although the growth curve is similar among mutants, fluorescence intensities varied among the mutants. In summary, it was found that substitution mutations occurring to the AGGAG sequence within BBa_R0034, AAAGAGGAGAAA, showed a decreased GFP expression while others showed increased GFP output. Especially for mutations to base pair 7, GFP expression is near total-depression.
After normalising the GFP fluorescence readings with the OD600 at T=8, the ratio of the normalised fluorescence of the mutants to wild type RBS is computed and plotted as shown.
We also spotted the mutants on an LB agar plate to have a qualitative view of the GFP brightness. The culture is diluted to OD600 = 0.4 then 3uL of the culture is spotted on to the agar. Brightness of these spots parallels the results in the above graph. For example, 7GA, 8AG and 9GA is the brightest among the mutations on their respective base pairs while 12AC and 11AC are the darkest.
