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==Usage Notes== | ==Usage Notes== | ||
A 1990 [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=2199796 paper] by [http://www.biology.bnl.gov/cellbio/studier.html Bill Studier] discusses how the secreted [[β-lactamase]] can quickly consume all the ampicillin in a culture (even at 20μg/ml Amp). A stationary culture of ampicillin resistant cells can have such a concentration of [[β-lactamase]] that even a 1/200 to 1/1000 dilution will still contain enough [[β-lactamase]] to consume all the fresh ampicillin before all the non-resistant cells from the stationary phase culture have been killed. | A 1990 [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=2199796 paper] by [http://www.biology.bnl.gov/cellbio/studier.html Bill Studier] discusses how the secreted [[β-lactamase]] can quickly consume all the ampicillin in a culture (even at 20μg/ml Amp). A stationary culture of ampicillin resistant cells can have such a concentration of [[β-lactamase]] that even a 1/200 to 1/1000 dilution will still contain enough [[β-lactamase]] to consume all the fresh ampicillin before all the non-resistant cells from the stationary phase culture have been killed. The authors reccommend not allowing cultures to reach stationary phase if you need a high proportion of cells to contain your plasmid. | ||
*[[Barry Canton]] & [[Matt Gethers]] have seen this occur when using the BioBrick plasmid, pSB1A3 expressing a high level of mCherry. | |||
[[Carbenicillin]] is much more resistant and would be preferred except for cost. Mixtures of ampicillin and carbenicillin are often used. | [[Carbenicillin]] is much more resistant and would be preferred except for cost. Mixtures of ampicillin and carbenicillin are often used. |
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