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outgrowth time andampicillin conc for low-copy numbe plasmid - (Mar/10/2011 )

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You should grow your cells at room temperature for extremely low-copy number plasmids. Otherwise, the growth rate of your cells will be higher than the copy rate of your plasmid, and you'll have problems. I would also use Carbenicillin instead of Ampicillin, it is more stable, and will reduce your background (and thus increase your yield) immensely.

-Rsm-

@Rsm

What kind of problems are you referring to?

-donny-

No growth of bacteria, or growth of bacteria without plasmid (to avoid that you should use Carbenicilin). It would seem like they do not survive Ampicillin or maintain the plasmid.

-Rsm-

Fortunately, no problem of such yet, even with 100ug/mL ampicillin. Plasmid yield has been reasonable with 10mL LB. So, we can simply replace ampicillin with carbenicilin for plasmids with Amp resistance?

-donny-

Yes, you can just replace it, also using the same concentrations (50 or 100mg/l). It is more stable at 37C, and thus your concentration of antibiotic will be maintained for longer time.

-Rsm-

i should try this..i'm new to plasmid work and having this problem too..

-pLASMID-
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