pCRblant

Frederik Börnke ricky_boernke at gmx.net
Fri Jan 26 02:14:20 EST 2001



"K.Fushimi" schrieb:
> 
> Hi all!
> 
> My friends have trouble in preparation of pCRblunt plasmid vector which
> contains the ccdB
> gene.  They store pCRblunt transformed E. coli with glycerol.
> But when they use the stock, they often get wrong plasmid.
> If you have some experience,  please give us advice!
> Which E. coli strain dose cause better or worse results?
> Can you store transformed E. coli with glycerol?
>

Hi Kazuo,

be sure your strain is a lac repressor overproducer like XL-I blue for
instance. It turns off ccdB expression pretty good and the plasmid can
easily be propagated in that strain. 
Why don't you just store the plasmid and retransform it whenever
necessary?

Cheers
Ricky



> Kazuo Fushimi, Ph.D., PostDoc
> National Institute of Neuroscience, NCNP, Japan(NIN)
> Address: 4-1-1 Ogawa-Higashi, Kodaira, Tokyo 187, Japan
> Telephone: 81-423-41-2711
> E-mail: fushimi at ncnp.go.jp
> 
> ---

-- 
*******************************************************************

Frederik Boernke
Research Group of  Molecular Plant Physiology
Institute for Plant Genetics and Crop Plant Research (IPK)
Corrensstr. 3
06466 Gatersleben
Tel.  039482 -5 321
Fax. 039482 -5 515
http://www.ipk-gatersleben.de






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