Problem to transform E. coli

Jay Hinton hinton at icrf.icnet.uk
Sun Feb 1 09:21:12 EST 1998


In article <2.2f.32.19971219090900.00679c48 at osiris.univ-rouen.fr>,
nathalie.porchet at UNIV-ROUEN.FR (U295) wrote:

> Dears netters,
> 
> I would like to transform pEGFP-N1 into E. coli (DH5-alpha) but it seems
> there is a problem. I use the technique decribed by Hanahan and Meselson. I
> would like to know if anybody has problems transforming GFP into E. coli
> with this technique. If anybody uses another technique, please mail me.
> 
> Sincerely yours,
> 
> Sebastien PARIS
> ___________________________________________
>                   
> INSERM U295
> Faculte de Medecine-Pharmacie de Rouen  
> Avenue de l'Universite                                
> B.P. 97                                                        
> 76803 St-Etienne-Rouvray Cedex
> France
> 
> Tel : (33)-02-35-65-52-74
> Fax : (33)-02-35-66-12-78
> E-Mail : nathalie.porchet at univ-rouen.fr
> ___________________________________________


Hi,

Your problem may be that EGFP can be toxic in E. coli in media containing
much NaCl. Brendan Cormack (who made the F64L S65T EGFP mutation) told me
about this. I always use LB media without NaCl, and have no problems
moving EGFP plasmids to E. coli.

Good luck,

      Jay



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