I Have a problem with E. coli GFP, I am culturing them in LB medium/ampicillin + IPTG, but not all the cells fluorescent after UV excitation (fluorescence microscope) although they grew in the presence of ampicillin.
Can anybody help please?
E. coli GFP I used is ATCC 25922GFP
An overnight culture from frozen stock in LB med with ampicillin at 200 rpm, 37 C. My working bacterial culture was: one in ten dilution of the overnight culture in LB medium with supplied with ampicillin and IPTG grew at thepreviuos condition for 2 hours.
Ampicillin concentration is 100 ug/ml.
IPTG was added only in the 2 H culture in a concentration of 0.2 mM.
Before that I examined the fluorescence intensity with spectrophotometer at various concentration of IPTG but there was no significant difference in the fluorescence intensity.
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