Best vector for cloning of Taq-generated PCR products

Deb Britt Deborah_Britt at brown.edu
Tue Sep 16 13:59:50 EST 1997


We've been using Novagen's "perfectly blunt" kit for PCR product cloning. 
For products generated with Taq polymerase, there is a 40 minute enzyme
treatment step, then ligation.  You can also clone products generated with
proofreading pols.  We've been quite satisfied with the system.

Deb Britt

In article <199709120613.XAA29485 at net.bio.net>, boernke at IPK-Gatersleben.de
(Frederik Boernke) wrote:

> Hi all!
> 
> We rountinely use pGEM-T for cloning of our Taq-generated PCR
> Products, but sometimes the effeciency seems to be not very reproduceable.
> May be this depends on several freeze thaw cycles. Also I think
> the MCS is not really satisfying, maybe at least some of the routinely
> used restrictionenzymes should cut.
> What are youre experiences, specially with the use of other T-tailed
> vectors.
> May be this has discussed here a trillion times, but why not do it again?
> 
> Ricky
> 
> ******************************************************************
> 
> Frederik Boernke
> Research Group of  Molecular Plant Physiologie
> Institute for Plant Genetics and Crop Plant Research (IPK)
> Corrensstr. 3
> 06466 Gatersleben
> Tel.  039482 -5 321
> Fax. 039482 -5 515
> e-mail: boernke at ipk-gatersleben.de
> http://www.ipk-gatersleben.de

-- 
Deborah Britt, Ph.D.
Department of Medical Oncology
Rhode Island Hospital




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