Description
Antarctic Phosphatase catalyzes the removal of 5´ phosphate from DNA and RNA. Since phosphatase-treated fragments lack the 5´ phosphoryl termini required by ligases, they cannot self-ligate. This property can be used to decrease the vector background in cloning strategies.
Abbr
Antarctic Phosphatase, Recombinant (E. coli)
Applications
Removing 5´ phosphates from DNA, RNA, rNTPs and dNTPs Preparation of templates for 5´ end labeling Prevention of recircularization of cloning vectors Removal of dNTPs and pyrophosphate from PCR reactions Dephosphorylation of proteins
Form
10 mM Tris-HCl (pH 7.4), 1 mM MgCl2, 0.01 mM ZnCl2, 1 mM DTT and 50% glycerol.
Molecular Weight
Apparent: 35 kDa Theoretical: 69 kDa
Purity
>95% estimated by SDS-PAGE
Concentration
5,000 units/ml
Unit Definition
One unit is defined as the amount of enzyme that will dephosphorylate 1 µg of pUC19 vector DNA cut with HindIII (5´ protruding ends), HincII (blunts ends) or PstI (5´ recessed ends) in 30 minutes at 37°C. Dephosphorylation is defined as > 95% inhibition of recirculation in a self-ligation reaction and is measured by transformation into E. coli.
Synonyms
Antarctic Phosphatase
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