Imperial/Wet Lab/Lab Notebook/2007-08-31
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#Transformed ligated plasmid into electrocompetent DH5alpha cells | #Transformed ligated plasmid into electrocompetent DH5alpha cells | ||
- | Protocols can be found at [ | + | 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 |
#Grew cells at 37°C for 30 min | #Grew cells at 37°C for 30 min |
Latest revision as of 12:59, 26 October 2007
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
- 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
- Grew cells at 37°C for 30 min
- Plated onto Kanamycin plates
- Left plates overnight at 37°C