John Richard Seavitt
jrseavit at artsci.wustl.edu
Tue Jul 22 14:21:00 EST 1997
On 22 Jul 1997, Woong Choi, M.D. wrote:
> Do anybody have an experience with E.coli strain MC1061/p3?
> I want to know about appropriate concentration of antibiotics
> [ampicillin, tetracycline, kanamycin] for MC1061/p3.
Invitrogen (who supplies MC1061/p3, btw) suggests 7.5-10ug/ml Tet and
25-40ng/mL Amp for selection of transformed bacteria, and 30ug/mL Kan for
maintenance of the P3 episome alone. I used only Kan for preparing the
empty vector for ligations (obviously) and only Amp/Tet for selection on
plates, etc. It's been suggested to me that the presence of Kan during
the preparation of supercompetant bugs decreases transformation effiency,
and this has been true in my hands in one side-by-side comparison.
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