31 August 2007

Cell by date - Operating Temperature Range (continue)

Construct - pTet-GFP [http://partsregistry.org/Part:BBa_I13522 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.

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 [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#S30.2FS12_Cell_Extract#Transformation| Transformation] in the general protocols page

1. Grew cells at 37°C for 30 min
2. Plated onto Kanamycin plates
3. Left plates overnight at 37°C