Antibiotics for M9 growth?
hzhen at freeuk.com
Sun Feb 20 17:01:45 EST 2005
> You don't really need 2 weeks to grow E.coli in deuterated media. I
> don't know what you want to do with the deuterated protein, but the
> one way is to grow in small volume (~5-10ml) in say 50% D2O media,
> spin down cells when ready, inoculated into 50-100 ml of 80-90% D2O
> media, then spin down and inoculate into 1L of 100% D2O. A few days
> is the most you need, and no problem with ampicillin/carbenicillin
> unless there is significant problem with toxicity of your protein. If
> you want to look for references for proper protocols, find papers by
> Kevin Gardner and Lewis Kay.
> Spinning down cells and inoculated into fresh media with fresh
> antibiotics is the way to maintain plasmid even if you want to grow
> the cells in 100% D2O media throughout. Start with small volume and
> increase volume in each step.
Forgot to mentioned, but you can also use deuterated rich media which
can help the cells to grow faster, a few companies sell those, such as
Silantes, Isotec and Spectra Stable Isotopes (formally Martek). If
you think that they are too expensive, spiking the minimal media with
a bit of the labelled rich media can help with the growth.
> > Thanks
> > kmj
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