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Advice in microbiology needed

Michael Witty mw132 at mole.bio.cam.ac.uk
Wed May 22 11:09:06 EST 2002


On 22 May 2002, Vince Mulholland wrote:

> Vladimir,
>
> Use chloramphenicol NOT ampicillin (the bacterium is Cam-R). This seems to
> be your problem.
>
> Hi,
>
> BL21 CodonPlus (DE3) RIL from Stratagene has genotype E. coli B F(-) ompT
> hsdS(rB(-) mB(-)) dcm(+) Tet(r) gal ?(DE3) endA Hte[argU ileY leuW Cam(r)],
> and appears it should be able to grow in a minimal medium. We tried to grow
> it in M9 with glucose (and ampicillin for plasmid maintenance) with no
> avail. Tech. rep. from Stratagene was not of much help suggesting to add
> proline (?), cysteine (?) and thiamine (?) with no guarantee to work. Any
> advice what may be a problem and how to make it grow?
>
> Thanks,
>
> Vladimir Yamshchikov


Vince, are you sure?  Vladimair, what about the idea of spreading a
minimal plate with cells and putting a grain or two of all the amino
acids on your chemical shelf in sectors and see what makes a difference
(classical experiment).  Regards, Mike.




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