Any experience with Hind3 on a primer?
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.
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 J. Witchel, Ph.D.
> Dept. Physiology
> Medical School
> Bristol BS8 1TD
> Harry.Witchel at Bristol.ac.uk
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