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Without any details regarding the transformation experiment, I can only provide some general comments based upon my laboratory experience:
1. If competent E. coli cells are transformed with a plasmid conferring ampicillin resistance, then incubation periods of greater than 24 hours is usually sufficient to recover ampicillin-resistant transformants. The same would also be true if the plasmid were introduced into E. coli by electroporation. If such an experiment repeatedly failed, then the plasmid may not be able to replicate in E. coli.
2. If ampicillin resistance is transferred to E. coli in a conjugation experiment (i.e., a mating experiment), then more than 24 hours may be needed for incubation in order to recover transconjugants. Some protocols recommend holding the plates for up to 5 days if the conjugation is predicted to be a low-frequency event.
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