TA cloning and competent cells

eric anderson e-anderson at ski.mskcc.org
Tue Jan 14 08:57:18 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

Ed,

chemical competent (CaCl2) DH5-alphaF' cells well in my hands.  you just
need to remember to use both X-Gal and IPTG if you want blue-white
screening to work.

good luck,

eric

-- 
Eric C. Anderson
Memorial Sloan-Kettering Cancer Center
Sloan-Kettering Institute
1275 York Ave. Box 470
New York, NY  10021
(212) 639-2977
e-anderson at ski.mskcc.org



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