3kB PCR Cycling Conditions?//

Anderas Becker becker at ps1515.chemie.uni-marburg.de
Thu Jul 20 03:06:52 EST 1995


In article <3ucuiv$imf at news.ycc.yale.edu>, burbly at minerva.cis.yale.edu (Marc A. Borbely) says:
>
>Hello. I'm trying to amplify a 3kB fragment from mouse genomic DNA. I've 
>been using Taq polymerase with a 3 minute extension time, and 20 seconds 
>annealing, and 20 seconds denaturing. I'm getting no product. Does anyone 
>have better cycling conditions I should try? 
>Thanks
>
>- Marc Borbely

Taq is not the best enzyme therefore. I prefer Vent-polymerase.
Did you check the template and the primers with a second PCR?
I did not have any problems wi
There are also "long PCR kits availible".

Andreas

----------------------------------------------------------------------
Andreas Becker
Arbeitskreis Prof. Kadenbach, FB Chemie/Biochemie, Hans-
Meerwein-Strasse, Philipps-Universitaet, 35043 Marburg, Germany
Phone: privat +49 6421 47304  Labor +49 6421 28 -5721 Fax -2191
eMail: BECKER at ps1515.Chemie.Uni-Marburg.De
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