TA cloning and competent cells

[Steven Goldberg]

In article <Pine.A41.3.95.970113183839.20104H-100000 at aix1.uottawa.ca>,
sunfish <s535290 at aix1.uottawa.ca> wrote:

> Hi all,
> 
> does anyone ever use homemade competent cells for use with TA cloning ? 
> (we use the pCR2.1 by invitrogen)  Sometimes we run out of the competent
> cells that come with the kit and if I'm in a hurry and can't wait for new
> cells to arrive, it'd be nice to know that the home made stuff is alright.
> If anyone does use their own competent cells, what strain do you use, and
> what method do you use for making the cells competent ?
> 
> thanks in advance,
> 
> Ed

I will at times have problems with the competent cells from Invitrogen
(either I run out of them or they seem to lose their competency).  In such
instances I substitute with home-made electrocompetent INValphaF' or
similar strain such as DH10B from BRL with no problems at all.

Steve