Alkaline Phosphatase, can remove the 5' phosphate groups from DNA and RNA. It is commonly used to prevent the self-ligation of vectors. In molecular cloning experiments, DNA ligase requires the presence of phosphate groups to catalyze DNA ligation. After digestion with restriction enzymes, vectors retain a phosphate group at the cleavage site. However, after dephosphorylation by Alkaline Phosphatase, the 5' end of the vector lacks a phosphate group and thus cannot connect with its own 3' end. As a result, the self-ligation of the vector, which would otherwise preferentially occur in the ligation reaction, is prevented. This increases the insertion rate of the target fragment. Additionally, Alkaline Phosphatase can be used to prepare DNA templates for 5' end labeling. This product is a shrimp-derived Alkaline Phosphatase that can degrade almost all phosphate monoesters. However, it cannot hydrolyze phosphate diesters or triesters. The enzyme acts on the terminal dephosphorylation, regardless of whether the termini are sticky or blunt. SAP can also be used to degrade dNTPs in PCR reactions, which is useful for subsequent sequencing and SNP analysis. Features Removal of phosphate groups from DNA and RNA ends. Preventing self-ligation of vectors. Preparation of DNA templates for 5' end labeling. Degradation of dNTPs in PCR reactions. Applications Molecular cloning DNA sequencing SNP analysis Components Components No. Name 10322ES72 10322-A Shrimp Alkaline Phosphatase (SAP) (1 U/µL) 250 U 10322-B 10×SAP buffer 1 mL Specifications Source From Pichia pastoris, the shrimp alkaline phosphatase gene was cloned from Pandalus borealis. Quality Control Free of ribonuclease and deoxyribonuclease contamination. Purity ＞95% Thermal Inactivation Incubation at 65℃ for 5 min results in complete and irreversible loss of activity. Unit definition The amount of enzyme required to dephosphorylate 1 μg of pUC19 DNA digested with HindIII (which produces 5' overhangs) within 30 min at 25℃ is defined as one unit of activity.
