In cloning, the vector is dephosphorylated by shrimp alkaline phosphatase and can not be linked to its 3' end because there is no phosphate group at the 5' end. Shrimp alkaline phosphatase can act on both 5' sticky ends and blunt ends, and can also be used to degrade free dNTP in PCR reaction for preparation of sequencing template and SNP analysis. The shrimp alkaline phosphatase can be completely and irreversibly inactivated by incubation at 65℃ for 5 min, so there is no need to remove it after ligation or end labeling. Based on above properties, this enzyme is an excellent alternative to the traditional dephosphorylase, calf intestinal alkaline phosphatase.
- Dephosphorylation of DNA and RNA
- Preventing self-ligation of cloning vectors
- Preparation of 5' end labeling template
- Removal of dNTP and pyrophosphate from PCR products
A unit is the amount of enzyme required to dephosphorylate 1 μg of pUC19 DNA digested by HindIII (producing a 5' protruding end) at 25℃ for 30 min. The dephosphorylation criterion refers to the inhibition of DNA reconnection by more than 95% in the self-ligation reaction.
Stored at -20℃ for 3 years.