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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