IGEM:IMPERIAL/2007/Experimental Design/Phase2/Results 2.2.1
Result Summary
pTet-LuxR-pLux-GFP
After testing 1nM and 1000nM we have now decided to test a range between 10nM,50nM and 100nM were tested for a period of 8 hours.
The results show us that as the AHL increase the out of fluorescence increases. In addition the response of the system was greater for a greater AHL concentration.
From these results we have now defined the range of interest as 1-10nM. Our initial specifications was to detect a threshold of 50nM, now however, we now our system is sensitive to at least 10nM. This makes our system a much more responsive and useful system for the infecter detector application.
From the results we also can see that for our system to reach steady state looks as if it takes 200-300 minutes. The specifications of the infecter detector system is to respond within 3 hours to give a visible output. We do not yet now if this will give a visible output with the final construct of dsRED.
Below are the averages of normalised data. The normalisation was done by taking into consideration the negative control, which was a cell extract with DNA of an empty plasmid. The reason why the data is initially negative is because the negative control has an initially high fluorescence level, compared to the others samples.
Complete set of results and raw data |