BL21(DE3) problems

[John T Jones]

Andy Scotter wrote:

>         We have just cloned a small insert (145bp) into pET42 and
> transformed BL21(DE3) cells with it.  They grow fine on plates containing
> LB, 1% Glucose and 15micrograms/ml Kanamycin LBGK), with a reasonable
> transformation efficiency.  However, I can't get a liquid culture growing
> using LBGK.  I've tried 2.5ml, 5ml and 10ml cultures in Universals  (i.e.
> plenty of air) and I'm incubating the culture at 37 degrees centigrade.
>         Anyone got any ideas?
>

You could try growing at 30.  This sometimes helps.  Dont remember the reasoning
off the top of my head I'm afraid.

John