<calendar> name=iGEM:IMPERIAL/2007/Notebook date=2007/09/24 view=threemonths format=%name/%year-%month-%day weekstart=7 </calendar>
Cell by date - Operating Temperature Range (continue)
Construct - pTet-GFP BBa_I13522
Temperatures - 37 °C and 20°C Aims: To test for the behaviour of the DNA construct (pTet) at temperatures 20oC and 37oC by observing the amount of fluorescence produced over a period of 24 hours.
- Measurements was carried out from the samples incubated overnight.
Preliminary AHL testing
Aim: This experiment was done to test the response time and steady state of the pTet-LuxR-pLux-GFP DNA construct at the different AHL concentrations, ranging from 100nM, 50nM, and 10nM.
GFP Calibration Curve
Aim: This experiment was done to generate a calibration curve of the amount of fluorescence generateed with respect to the concentration of GFP in the solution.
Construction of pT7-GFP
- Transformed ligated plasmid into electrocompetent DH5alpha cells
Protocols can be found at Transformation in the general protocols page
- Grew cells at 37°C for 30 min
- Plated onto Kanamycin plates
- Left plates overnight at 37°C