Transformation problems...

ChenHA hachen at bun.com
Mon Jan 17 08:38:26 EST 2000


khalil wrote:

> All possible methods are being used, we all work in different labs in the
> departement.
> 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
> would
> > 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





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