Difference between revisions of "Team:Newcastle/Proof/VariableResistor"
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<p> The cells were grown up in 10ml of liquid culture of LB broth with chloramphenicol overnight at 37°C. The cells were then diluted down to an OD of 0.05 at 600nm and 100μl of the cells were then added to each well. The cells were then left to grow for 16 hours and the OD600 was measured every 5 minutes. The results can be seen below in figure 2. </p> | <p> The cells were grown up in 10ml of liquid culture of LB broth with chloramphenicol overnight at 37°C. The cells were then diluted down to an OD of 0.05 at 600nm and 100μl of the cells were then added to each well. The cells were then left to grow for 16 hours and the OD600 was measured every 5 minutes. The results can be seen below in figure 2. </p> | ||
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| + | <IMG SRC="https://static.igem.org/mediawiki/2016/0/01/T--Newcastle--VR_with_Arabinose.png" ALT="some text" WIDTH=32 HEIGHT=32> | ||
<i> Conclusion </i> | <i> Conclusion </i> | ||
<p>From our experiments, we can conclude that our ‘Variable Resistor’ (VR) construct does not grow in ZnSO<sub>4</sub> when arabinose is present. The cells only grew when zinc was not present. Therefore, showing that our construct doesn’t sequester zinc when the SmtA protein is produced. It is possible that this is because SPEAKING TO JON ON MONDAY to give possible reasons about this. </p> | <p>From our experiments, we can conclude that our ‘Variable Resistor’ (VR) construct does not grow in ZnSO<sub>4</sub> when arabinose is present. The cells only grew when zinc was not present. Therefore, showing that our construct doesn’t sequester zinc when the SmtA protein is produced. It is possible that this is because SPEAKING TO JON ON MONDAY to give possible reasons about this. </p> | ||
Revision as of 18:42, 16 October 2016
Results
Growth in Varying ZnSO4 Concentrations (Minimum Inhibitory Concentration):
In this experiment, we used 6mM of zinc sulphate (ZnSO4) and carried out a dilution of half for each column of a 96 well plate to try and identify the minimum inhibitory concentration for the E. coli to grow in. Cells were again placed in the pSB1C3 backbone and fresh cultures were grown at 37°C overnight. The cells were diluted down to a starting optical density of around 0.05 at 600nm using LB broth with chloramphenicol. Then, 100μl of cells were placed in the wells with the corresponding ZnSO4 concentration which was diluted using LB with chloramphenicol. This experiment was left for 16 hours to give the E. coli time to grow and replicate, see figure 1.
Figure 1.
From this experiment, we found that the minimum inhibitory concentration was 3mM of ZnSO4 for E. coli. This concentration was then used in our second experiment to test whether our “Variable Resistor” (BBa_K1895999) construct was able to sequester zinc in the presence of arabinose.
Growth with Zn and Arabinose
The minimum inhibitory concentration was used for this experiment to determine whether the construct would sequester the zinc if arabinose was present. For this, we used a 96 well plate and used half of the plate to contain 3mM of ZnSO4 with ???mM of arabinose and LB broth with chloramphenicol. The other half of the plate contained the ???mM of arabinose with LB and chloramphenicol.
The cells were grown up in 10ml of liquid culture of LB broth with chloramphenicol overnight at 37°C. The cells were then diluted down to an OD of 0.05 at 600nm and 100μl of the cells were then added to each well. The cells were then left to grow for 16 hours and the OD600 was measured every 5 minutes. The results can be seen below in figure 2.
<IMG SRC="
" ALT="some text" WIDTH=32 HEIGHT=32>
Conclusion
From our experiments, we can conclude that our ‘Variable Resistor’ (VR) construct does not grow in ZnSO4 when arabinose is present. The cells only grew when zinc was not present. Therefore, showing that our construct doesn’t sequester zinc when the SmtA protein is produced. It is possible that this is because SPEAKING TO JON ON MONDAY to give possible reasons about this.
