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Amp selection with no recovery

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Amp selection with no recovery

Postby daknecht » Sat Dec 08, 2012 4:54 pm

For years, I have been doing E. coli transformation by heat shocking cells and then plating immediately on amp plates with no recovery. I was taught this trick years ago and was told that the reason it worked had to do with the fact that amp affects cell wall biosynthesis (rather than translation) and once the cells began making ampicillinase, their cell walls could recover and cells would grow. Translational inhibitors obviously would not work with this scheme as you would never make the resistance gene without recovery.
My question is that I cannot remember who told me this and I have been unable to find the source of this information. In fact, every protocol I can find online uses a 30-60 min recovery period following heat shock. Does anyone know a reference that documents this information or refutes it?
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Postby JackBean » Mon Dec 10, 2012 5:52 pm

if it works for you, then it should be right ;) and it sounds reasonable

Cis or trans? That's what matters.
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