Antarctic Phosphatase (AnP) is a heat labile alkaline phosphatase purified from a recombinant source.
- Rapid and irreversible heat inactivation eliminates unwanted activity
- Flexible reaction conditions (active in any restriction enzyme buffer, no clean-up required)
- No need for multiple phosphatases (AnP removes 5´- and 3´- phosphates from DNA, RNA and dNTPs)
- Active on unincorporated dNTPs in PCR products - improves DNA sequencing and SNP analysis
Antarctic Phosphatase (AnP) is a heat labile alkaline phosphatase purified from a recombinant source. AnP nonspecifically catalyzes the dephosphorylation of 5´ and 3´ ends of DNA and RNA phosphomonoesters. Also, AnP Hydrolyses ribo-, as well as deoxyribonucleoside triphosphates (NTPs and dNTPs). AnP is useful in many molecular biology applications such as the removal of phosphorylated ends of DNA and RNA for subsequent use in cloning or end-labeling of probes. In cloning, dephosphorylation prevents religation of linearized plasmid DNA. The enzyme acts on 5´ protruding, 5´ recessed, and blunt ends. AnP may also be used to degrade unincorporated dNTPs in PCR reactions to prepare templates for DNA sequencing or SNP analysis. AnP is completely and irreversibly inactivated by heating at 80°C for 2 minutes, thereby making removal of AnP prior to ligation or end-labeling unnecessary.
An E. coli strain that carries the TAB5 AP gene, originally cloned in plasmid pNI (2), recloned in plasmid pEGTAB7-4.1(3).
- Dephosphorylation 5´ and 3´ ends of DNA and RNA.
- Dephosphorylation of cloning vector DNA to prevent recircularization during ligation.
- Dephosphorylation of DNA prior to end-labeling using T4 Polynucleotide Kinase.
- Treatment of dNTPs in PCR reactions prior to sequencing or SNP analysis.