IGEM:IMPERIAL/2007/Notebook/2007-8-31

name=iGEM:IMPERIAL/2007/Notebook date=2007/09/24 view=threemonths format=%name/%year-%month-%day weekstart=7

Cell by date - Operating Temperature Range (continue)
Construct - pTet-GFP BBa_I13522 Temperatures - 37 &deg;C and 20&deg;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.

Protocol can be found here under Phase 2-Operating Temperature Range on the experimental design page. Results can here under Results on the experimental design page.

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.

Protocol can be found here under Phase 2-Preliminary AHL testing on the experimental design page. Results can here under Results on the experimental design page.

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.

Protocol can be found here under Phase 2-GFP calibration curve on the experimental design page. Results can here under Results on the experimental design page.

Construction of pT7-GFP

 * 1) Transformed ligated plasmid into electrocompetent DH5alpha cells

Protocols can be found at Transformation in the general protocols page


 * 1) Grew cells at 37&deg;C for 30 min
 * 2) Plated onto Kanamycin plates
 * 3) Left plates overnight at 37&deg;C