The amplitude of change between GFP expression at 0 AHL and 10uM AHL seems to a lot smaller in comparison to the experiment on 8/8/06.
Flourescence decreased over the time period
This suggests the peak of GFP expression is even earlier than 3h
This could either be down to
A very short AHL Half-Life
Cell death caused by poor heat conductance and lack of shaking in the water bath (used after the first fluorescence measurement, no shaker in plate reader room!)
Conclusions
Repeat experiment
Try and find a shaker
Experimentally assess AHL half-life
Well Layout
14/8/06
Aims
To run through T9002 protocol and achieve transfer function plot
Extra Information
Due to a protocol error the cultures could not be innoculated with AHL at OD600s of 0.1. Instead they were innoculated at the following ODs:
T9002 LB: 0.05
T9002 M9: 0.03
The protocol error has since been corrected.
Results
Graphs
Average T9002 response to AHL in LB over time periods 4h, 4h20, 4h40, 5h (Line)
Average T9002 response to AHL in M9 over time periods 4h, 4h20, 4h40, 5h (Line)
The results show that the cultures in M9 had a higher amplitude of fluorescence
The results seem to suggest that the cultures in M9 would be responsive to greater concentrations of AHL than those tried, in contrast to the experiment on the 8th, and MIT's experiments
This could be due to the innoculation OD600
Unsure on how to remove media background fluorescence
Levels of GFP generally went down over the measured time period
This suggests GFP had already reached it's peak of expression at an earlier time
It also suggests AHL half-life may be shorter than the assumed 24 hours
Conclusions
The experiment should be repeated at OD600 of 0.1
A way to mathmatically remove background flourescence should be devised
AHL half-life should be experimentally measured
Well Layout
8/8/06
Aims
This experiment was a basic test run of the T9002 protocol. It was also designed to give insight into whether to use LB or M9 media for our characterisations and, eventually, full system.