Bacterial Alkaline Phosphatase

Bacterial Alkaline Phosphatase (BAP) removes 3´ and 5´ phosphates from DNA and RNA. BAP is active at 65°C for at least 1 h and is inactivated by phenol extraction.

Applications: Dephosphorylation of 5´-phosphorylated termini of vector DNA to prevent self-ligation. Dephosphorylation of 5´ termini of nucleic acids prior to forward reaction with kinase.

Source: Purified from E. coli C90.

Performance and Quality Testing: Endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; dephosphorylation
efficiency determined.

Unit Definition: One unit hydrolyzes 1 nmol of ATP in 30 min. at 37°C.

Unit Reaction Conditions: 10 mM Tris-HCl (pH 8.0), 1.5 mM [γ-³²P]ATP, and enzyme in 50 μl for 30 min. at 37°C.