Bacterial alkaline phosphatase (BAP) removes 3´ and 5´ phosphates from DNA and RNA. BAP remains active at 65° C for at least 1 hour and is inactivated by phenol extraction.

Application:Dephosphorylation of the 5´-phosphorylated end of vector DNA to prevent self-ligation. Dephosphorylation of the 5´ end of nucleic acids before the forward reaction of the kinase.

Source: Purified from E. coli C90.

Performance and quality testing: endodeoxyribonuclease, exodeoxyribonuclease and ribonuclease detection; determination of dephosphorylation
efficiency.

Unit definition: At 37°C, one unit hydrolyzes 1 nmol of ATP in 30 minutes.

Unit reaction conditions:10mM Tris-HCl (pH 8.0), 1.5 mM [γ-³²P]ATP and enzyme, 50 μl, after 30 minutes at 37°C.