Imperial/Wet Lab/Lab Notebook/2007-08-31

From 2007.igem.org

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==Cell by date - Operating Temperature Range (continue)==
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'''Construct - pTet-GFP [http://partsregistry.org/Part:BBa_I13522 BBa_I13522]'''<br>
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'''Temperatures - 37 &deg;C and 20&deg;C'''
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'''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. <br>
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*Measurements was carried out from the samples incubated overnight.
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Protocol can be found here under [[IGEM:IMPERIAL/2007/Experimental_Design/Phase2/Protocol_2.1.1|Phase 2-Operating Temperature Range]] on the experimental design page.
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<br><br>
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Results can here under [[IGEM:IMPERIAL/2007/Experimental_Design/Phase2/Results_2.1.1| Results]] on the experimental design page.
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==Preliminary AHL testing==
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'''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.
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Protocol can be found here under
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[[IGEM:IMPERIAL/2007/Experimental_Design/Phase2/Protocol_2.2.1|Phase 2-Preliminary AHL testing]] on the experimental design page.
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<br><br>
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Results can here under [[IGEM:IMPERIAL/2007/Experimental_Design/Phase2/Results_2.2.1| Results]] on the experimental design page.
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==GFP Calibration Curve==
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'''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.
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Protocol can be found here under
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[[IGEM:IMPERIAL/2007/Experimental_Design/Phase2/Protocol_1.2|Phase 2-GFP calibration curve]] on the experimental design page.
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<br><br>
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Results can here under [[IGEM:IMPERIAL/2007/Experimental_Design/Phase2/Results_1.2| Results]] on the experimental design page.
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==Construction of pT7-GFP==
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#Transformed ligated plasmid into electrocompetent DH5alpha cells
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Protocols can be found at [[IGEM:IMPERIAL/2007/Notebook/General_Protocols#Transformation|Transformation]] in the general protocols page
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#Grew cells at 37&deg;C for 30 min
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#Plated onto Kanamycin plates
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#Left plates overnight at 37&deg;C
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Revision as of 14:05, 10 October 2007

Contents

31 August 2007

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.

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

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