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+ | Description : | ||
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+ | We used plasmids containing different promoter from the 2017 distribution kit and transfect into E.coli. Before measuring fluorescence expression, we used LUDOX and fluorescein to calibrate the TECAN. | ||
+ | We prepared a dilution series of fluorescein in 4 replicates and measure the fluorescence in a 96 well plate in a plate reader. By measuring these in all standard modes,we generated a standard curve of fluorescence for fluorescein concentration.We used this to correct cell based readings to an equivalent fluorescein concentration and convert this into a concentration of GFP. |
Revision as of 13:01, 18 August 2017
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Description :
We used plasmids containing different promoter from the 2017 distribution kit and transfect into E.coli. Before measuring fluorescence expression, we used LUDOX and fluorescein to calibrate the TECAN. We prepared a dilution series of fluorescein in 4 replicates and measure the fluorescence in a 96 well plate in a plate reader. By measuring these in all standard modes,we generated a standard curve of fluorescence for fluorescein concentration.We used this to correct cell based readings to an equivalent fluorescein concentration and convert this into a concentration of GFP.