DH5a in DMEM
karaolis at umabnet.ab.umd.edu
Mon Feb 19 21:36:39 EST 1996
On 19 Feb 1996, Chris Boyd wrote:
> David Karaolis (karaolis at umabnet.ab.umd.edu) wrote:
> : I have a cosmid clone in DH5a and want it to grow in DMEM (tiss cult med).
> : I can't get this strain to grow at all in standard DMEM.
> Why do you want to do this? Most folk are busting their guts
> trying to STOP bacteria growing in DMEM.
Reason- I want to do this as it has been shown certain proteins are
secreted in the presence of this medium.
> : I know the cosmid is not toxic etc as this strain grows well in
> : standard LB broth.
> : Does anyone know how to get it to grow in DMEM (additives etc?)
> As another person has said, you should check if antibiotics have been
> added. Also, why not grow the bacteria to exponential phase in LB,
> wash and resuspend in DMEM (minus antibiotics of course)?
Reply- I am using standard MEM without antibiotics.
The suggestion of growing first to log in LB, washing then inoculating MEM
is a good one. I have inoculated MEM from an overnight plate and this did
not work. Maybe a few hours first in LB may work.
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