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excessive ampicillin in L.B/Amp plates - (Jul/16/2008 )

I accidently add excessive ampicillin to L.B and used that for making agar plates ( 800ul instead of 400ul in 400ml of L.cool.gif. I need to used them for selection of HD alpha 5/pUC19, would it be a problem. Can I use these plates or should I make new ones ?
Thank you

-David646-

They will probably work. pUC19 is a high copy plasmid, and the amp resistance gene conveys high resistance. I'd definitely try them. You might want to run a control of plating out a known resistant strain to check that it grows, so that you will know if the plates are a problem, in the event of difficulties.

-phage434-

Either you can try it or just leave it for weeks and the amp will start to degrade. And then you can start to use them. but I am always very skeptical about the concentration. wink.gif

-timjim-

QUOTE (David646 @ Jul 17 2008, 03:51 AM)
I accidently add excessive ampicillin to L.B and used that for making agar plates ( 800ul instead of 400ul in 400ml of L.cool.gif. I need to used them for selection of HD alpha 5/pUC19, would it be a problem. Can I use these plates or should I make new ones ?
Thank you


So you doubled? But anyway, what is your end-concentration?

However, I do agree with phage434.

-Pallas-

Thank you for reply, I am going to used them I hate waste.

-David646-