primers for TA cloning

Vijay Sarthy vjsarthy at merle.acns.nwu.edu.
Wed Nov 15 18:32:17 EST 1995


In article <celer.4.0007199E at brno.ics.muni.cz>, celer at brno.ics.muni.cz wrote:

> Hi,
> I can not find the information about first nucleotid in my primers which will
> be used in TA cloning. I heard, that termostable polymerase is addind an A as
> overhang depending of the first (or last) nucleotid in primer. Can you send me
> this information?
> Celer Vladimir
> Veterinary univerzity Brno  Czech rep

Taq polymerase adds a 3' A independently of the PCR product you have
amplified. The primers used make no difference, but be sure to ligate to
the TA vector immediately following amplification to avoid degradation of
the A overhangs.

good luck,
Bill Cook
Northwestern



More information about the Methods mailing list