Cells with pET16b are not growing in liquid Medium

Richard J. Dudley rjdudley at YAHOO.COM
Wed Aug 12 10:11:15 EST 1998


---Matthias Drosten <Matthias.drosten at rz.ruhr-uni-bochum.de> wrote:
>
> 		
> Hello,
> I have got the following problem:
> 
> After transforming a ligation with pET16b and an insert into XL1 blue
> I get about 60 colonies (7 colonies at the religation control) on TBY
> + Amp.   The grew very slowly.  Transferring them into liquid medium
> they are not growing at all (at the moment nothing had happened for  2
> Days.). Transferring them to an agarplate they are growing again.
> 
> Has anyone an idea to solve this Problem?? I want to isolate the
> plasmide to verify the ligation.
> 
> Thanks for help
> 
> Heiko
> 
> Heiko.Koch at ruhr-uni-bochum.de

1) Repeat the same transformation-plating-liquid culture regimen with
the parent vector and one of the colonies form the religaiton control.
2) What is the [amp] in liquid culture?
3) Check your plates.
4) Call Novagen, or whomever you got the vector from, for suggestions
on using this vector with a toxic insert.
5) Search the bionet archives (http://www.bio.net) for info on growing
a toxic insert.

In my experiences, I've seen about 50-50 technique vs. something weird
and inexplicable happening.

rich
==
--- --- --- -- -- -- --- --- ---
Richard J. Dudley (rdudley at iname.com)
Research Specialist V
Dept. of Cell Biology and Physiology
The University of Pittsburgh
->biosci archives can be searched at www.bio.net <-

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