> All possible methods are being used, we all work in different labs in the
> In ours we use the CaCl2 method and a new method involving TSS. Competent
> cells are different for each one and stored in different freezers. We don't
> even use the same strain of E.coli.
I think what we need to know is the controls you made. For example, what is
the transformation efficiency? I cannot believe that you don't get any
colonies using uncut plasmids, so I presume you mean you don't get colonies
from ligation reaction. Is therefore the ligation reaction to blame? Did you
use commercially prepared cells as a control? etc. etc.
>> "Kiley R. Prilliman" <tiey at aol.comjunkbloc> wrote in message
> news:20000117030214.10870.00000733 at ng-fg1.aol.com...> > Why don't you tell us a little about the method you all are using? That
> > help us out in addressing any problem(s)...
> > best,
> > Kiley
> > Kiley R. Prilliman, Ph.D.
> > Postdoctoral Research Fellow
> > Division of Immune Regulation
> > La Jolla Institute for Allergy & Immunology
> > kiley at liai.org