Any experience with Hind3 on a primer?

Zhang Zhiying zhangz at ERE.UMontreal.CA
Sun Dec 7 12:38:00 EST 1997


Hi Harry Witchel;
I used Hind3 on a PCR primer for directional cloning and it works fine. 
When adding Hind3 site to the 5' end of a primer, it is improtant to add 
3 bases (I used GCG) to serve as a clamp to keep the 5' end from 
"breathing" during digestion.
good luck.

On Sun, 7 Dec 1997, Harry Witchel wrote:

> Date: Sun, 7 Dec 1997 15:43:15 GMT
> From: Harry Witchel <Harry.Witchel at Bristol.ac.Uk>
> To: methods at net.bio.net
> Subject: Any experience with Hind3 on a primer?
> 
> Hi folks --
>     Anyone got any experience using Hind3 on a PCR primer for
> directional cloning?  I would very much like to use Hind3, but according
> to the appendix of the New England Biolabs catalogue ("Cleavage close to
> the end of DNA fragments") Hind3 is quite inefficient cutting near the
> ends of DNA (eg at the 5' end of a PCR product).
>     Thanks much,
>         Harry
> 
> --
> Harry J. Witchel, Ph.D.
> Dept. Physiology
> Medical School
> Bristol  BS8 1TD
>    England
> 
> Harry.Witchel at Bristol.ac.uk
> http://www.bris.ac.uk/Depts/Physiology/Staff/hw.htm
> 
> 
> 



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