competent cells advice?
dk at no.email.thankstospam.net
Sat Nov 10 22:30:10 EST 2001
ms2241 at columbia.edu (Melanie Sohocki) wrote:
> I am having some insert instability problems with my subcloning.
>In the past, I have used the SURE cells from Stratagene to combat this
>problem, but both strains of these cells are resistant to kanamycin, the
>same marker on my plasmid. Unfortunately, I cannot simply change vector,
>because it is for a yeast-two hybrid application downstream. Any advice
>would be appreciated!
Just an idea: Avoid liquid cultures (o/n cultures in particular) by all
means. All problems with insert instability/toxicity are greatly reduced
when one tries to do as much as possible on plates and as little as possible
Don't ask me why because I don't quit understand it, but I know it
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