Ampicillin plates

Flip Hoedemaeker fhoedem at oci.utoronto.ca
Wed Dec 13 15:11:45 EST 1995


Dr. Arthur L. Kruckeberg wrote:
> 
> In article <gaughan-1112951612290001 at gaughangmac.wf.bms.com>
> gaughan at bms.com (Glen Gaughan) writes:
> 
> We use carbenicillin sodium B.P. (trade name is Pyopen) from Link
> Pharmaceuticals Ltd., Horsham, Surrey. CR7 7EQ. England. This 
appears
> to be more stable than ampicillin. We resuspend in sterile H2O at
> 100mg/ml and store in 500ul aliquots @ -20C. LB-carb (50-100ug/ml) 
agar
> plates appear to be stable for upto four weeks before satellite
> colonise appear after O/N incubation @37C. Plates are stored @ 4C 
and
> allowed to warm up to RT on the bench before cells are 
streaked/spread.
> 
> Hope it helps.
> 
> Chris

I've used Cb before, it seemed that especially with high copy number
plasmids (pETs pUCs etc.) you need higher amounts to prevent 
satellites (up to 250 ug/ml!) It is much more stable than Amp, but 
also much more expensive.

I'm in another lab now, and switched back to Amp. Making fresh plates
when you need them is the best solution to prevent satellites.
Just make plain plates, and streak them with Amp stock, them allow it 
to diffuse in for 5 minutes before you plate your cells. 
It also easier when you need a lot of different antibiotic 
combinations. (+/- pLysS for example....) 
Works for me! 

Flip



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