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Revision as of 16:10, 17 October 2018
Protocols
Lysing cells and using PCAU
PcaU fluorescence to measure [PCA]
-Culture BL21 E. coli containing PcaU fluorescent reporter in 50mL flask, 30C, 250rpm
-Prepare multi well plate containing serial dilutions of concentrated PCA solution in wells. Fill gaps between wells with water to prevent drying. Fill wells on edges with water.
-When OD600 = 0.6-0.8, transfer a fixed volume of culture from the flask to each well except for the water wells on edges
-Incubate >6 hours
- Read fluorescence in plate reader at 395 excitation 509 emission
- Optional: spin down plate and resuspend wells in PBS for reduced error
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