TA cloning and competent cells

David Micklem seemysignature at mole.bio.cam.ac.-uk
Tue Jan 14 08:38:58 EST 1997


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

You can afford to _buy_ competent cells routinely???!

The Inoue method of chemical competent cells is excellent and works with
every strain that I've tried - OK, so thats only XL1-blue and DH5alpha. 
Why don't you streak out some of the cells that come with the kit, and use
them? Make a glycerol stock while you are at it, so that next time you need
them you have a nice stock of the _original_ isolate.

I think the Inoue reference is in the FAQ.  Its in Gene, 1990 I think - I
can check if you can't find it.

David

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