Products > Molecular Biology > RNA & DNA Modifying Enzymes > DNA Manipulation

Antarctic Phosphatase (RK20537)

Datasheet

Summary

Catalog No.: RK20537

Concentration: 5, 000 U/ml

Background

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.

Product Component

Antarctic Phosphatase (5, 000 U/ml)RM21308
10X Antarctic Phosphatase Reaction BufferRM20152

Product information

Product SourceAn E.coli strain that carries the T4 DNA ligase gene.
ApplicationsDephosphorylation, RNA Modification, In vitro Synthesis (IVT)
Unit DefinitionOne unit is defined as the amount of enzyme that will dephosphorylate 1 ug of pUC19 vector DNA cut with a restriction enzyme generating 5' recessed ends in 30 minutes at 37°C. Dephosphorylation is defined as > 95% inhibition of recircularization in a self-ligation reaction and is measured by transformation into E. coli.
Storage Conditions10 mM Tris-HCl, 1 mM MgCl2, 50% Glycerol, 0.01 mM ZnCl2, pH 7.4 @ 25°C
Storage Temperature-20°C
Reaction Conditions1X Antarctic Phosphatase Reaction Buffer, Incubate at 37°C
Reaction Buffer or Mix1X Antarctic Phosphatase Reaction Buffer:
50 mM Bis-Tris-Propane-HCl, 1 mM MgCl2, 0.1 mM ZnCl2, pH 6 @ 25°C
Heat Inactivation80°C for 2 min

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