get rid of Alkaline phosphatase

Mike Cooley cooley at waksman.rutgers.edu
Sat Sep 11 22:35:56 EST 1999


"Kajetan H. Groicher" wrote:

> Sue-
>
> I routinely use Shrimp Alkaline Phosphatase rather than Calf Intestinal
> Alk Phos.  SAP is easily and irreversibly inactivated by heating at 65oC
> for ~30min.  CIAP is resistant to  heat inactivation.  I have found that
> SAP works as well as CIAP in my hands.
>
> The SAP dephosphorylated vector can be directly ligated to your insert
> without the need for further purification.
>
> Control transformations consisting of dephosphorylated vector treated
> with Ligase etc. has consistently yielded <<1% of my dpVector + Insert.
>
> Hope this helps.
>
> Kajetan

Where did you get your enzyme? We had terrible problems with SAP from
Amersham.


--
  ------------------------------------------------------------------------

Dr. Michael Cooley
Waksman Institute
Rutgers University
190 Frelinghuysen Rd.
Piscataway, NJ 08854

732-445-3802
FAX 732-445-5735
cooley at waksman.rutgers.edu




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