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.
For Research Use Only. Not for use in diagnostic procedures.
Contents & storage
Bacterial Alkaline Phosphatase is supplied with a vial of 10X dephosphorylation buffer [100 mM Tris-HCl (pH 8.0)]. Store at -20°C.