We collaborated with the Newcastle Team to help characterise one of their parts, the deGFP construct, which acts as a reporter in their improved biosensor framework called Sensynova. We offered to use a flow cytometer to thoroughly quantify the expression of the deGFP construct, using fluorescence-activated cell sorting (FACS) as this method was not accessible for the Newcastle iGEM team.
After the arrival of the DNA, we transformed the deGFP construct into E. coli Top10 cells before we ran the sample through the flow cytometer. This would sort from a stream of bacterial culture individual cells based on their properties, specifically fluorescence. Culture fluorescence was excited at 488 nm and intensity was recorded using a 530/30 nm detector to obtain the mean, median and standard deviation of a sample of 10,000 cells from the original population. We have sent both the raw and analysed data to Newcastle for their further analysis and interpretation.
In return Newcastle conducted a repeat of our experiment into testing the ability of UV to kill bacteria. By carrying out this experiment it meant that we could determine whether the results we obtained were reproducible, follow this link to see the results.
We also collaborated with the University of Cardiff iGEM team who conducted further experiments into testing the effectiveness of UV for the purpose of destroying bacteria. Due to time constraints we could not carry out these experiments ourselves. We felt that it was important to extend the experiment to see how the percentage of bacteria that had survived irradiation changed with a greater amount of exposure for instance, 20 minutes or 30 minutes. Additionally by collaborating with another team it meant that we could guarantee the reproducibility our of results. We sent a plate containing Top 10 E.coli to Cardiff and then using the protocol we had devised they irradiated 1 in 100,000 , 1 in 10,000,000 and 1 in 100,000,000 sample solution for five different exposure times. Most important Cardiff conducted the experiment for the extreme exposure times 1 minutes, 20 minutes 30 minutes, which we were not able to test ourselves.To see the result Team Cardiff obtained for us, click here.