get rid of Alkaline phosphatase

Kajetan H. Groicher Kajetan-Groicher at ouhsc.edu
Wed Sep 8 13:11:42 EST 1999


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
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