IGEM:IMPERIAL/2007/Experimental Design/Phase2/Results 2.1.1: Difference between revisions
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==Discussion== | |||
The results show that the fluorescence produced by the pTet-GFP construct is quite a linear function of time and it increases with increasing temperature. For our applications a visual signal is very important but even with the higest fluorescence, no fluorescence could be visualied even with a UV light. | |||
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'''31-08-2007''' | '''31-08-2007''' | ||
Revision as of 09:25, 17 September 2007
Result Summary
pTet-GFP
Aim
The aim of these experiments was to firgure out the operating range of the pTet-GFP construct in-vitro. The temperatures the construct was tested in are: 8°C, 10°C, 15°C, 20°C, 25°C, 30°C, 37°C.
Results
14-09-2007
The following graphs show how fluorescence produced by the pTet-GFP construct varied, on average, over time, as compared to the negative control, for different temperatures.
8°C
The fluorescence produced by the construct increases quite rapidly over the 6 hour period. But testing with UV light revealed that the fluorescence doesn't reach the visual threshold even after 6 hours.
| Complete set of results and raw data |
15°C
The construct seems to produce negligible fluorescence for the first 30mins of the reaction, if the negative control is taken into account. The maximum fluorescence is about 4 times more than that reached when the temperature is 8°C. The fluorescence was not visible at this temperature too, after 6 hours of reaction.
| Complete set of results and raw data |
25°C
The maximum fluorescence reached by the pTet-GFP construct at 25°C is approximately 4 times greater than that reached at 15°C. The highest fluorescence is about 160000, but even that was not visible with UV light.
| Complete set of results and raw data |
30°C
| Complete set of results and raw data |
Discussion
The results show that the fluorescence produced by the pTet-GFP construct is quite a linear function of time and it increases with increasing temperature. For our applications a visual signal is very important but even with the higest fluorescence, no fluorescence could be visualied even with a UV light.
31-08-2007
10°C
pTet was tested at 10oC for 4 hours. The results show no expression of pTet-GFP. After this experiment we took the samples out and put them at room temperature over night, this is to see if the low temperature permanently inactivate the in vitro chassis or DNA.
| Complete set of results and raw data |
20°C
pTet was tested at 20°C for two days.
| Complete set of results and raw data |
37°C
pTet was tested at 37°C for two days and a staggered experiment was carried out to minimise the missed measurements over night.
| Complete set of results and raw data |
