Low concentration miniprep
Aawara Chowdhury
via methods%40net.bio.net
(by aawara from pontiff-playground.org)
Sun Jan 25 10:37:26 EST 2009
In <6u1gnhFcaahfU2 from mid.individual.net>,
Christian Praetorius <prae from gmx.net> wrote:
> Duncan Clark <blackhole from abuse.plus.com> wrote:
>
>>The use of more beta lactamase resistant antibiotics such as
>>carbenicillin will reduce satellite formation. I still use 80:20
>>Methicillin:ampicillin, as I have a very old large bottle of methicillin
>>from before Sigma stopped selling it.
>
> Are cells carrying a ampicillin resistence plasmid resistent to
> carbenicillin/methicillin?
DH5a carrying pBR322/pMB15-based low-copy plasmids are resistant to
carbenicillin (at 100 ug/ml). I prefer carbenicillin to ampicillin
because it is more resistant to degradation by secreted beta-lactamase,
and significantly reduces the number of satellite colonies. Also, our
carbenicillin plates stay good in the cold room for over 6 months.
We have also used Timentin for the same purpose (it is a mixture of
a beta-lactam antibiotic - ticarcillin, and the beta-lactamase i
nhibitor clavulonic acid). Somewhat difficult to obtain without a
prescription these days.
AC
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