DNA modification enzymes

A high specific activity, heat-labile alkaline phosphatase. Completely and irreversibly inactivated in Tris buffers at pH 8,0 to 8,5 by heating for 15 minutes at 65 °C. No further treatment is necessary.

• Bacterial alkaline phosphatase recombinantly expressed in E coli
• Active in PCR and restriction endonuclease buffers
• Inactivated by heating at 75°C for 5 minutes
• Easily soluble in cold water and hot water
• Sourced from arctic shrimp (Pandalus borealis)

Use together with illustra Exonuclease I for cleaning up PCR reactions prior to downstream application such as DNA sequencing. Dephosphorylation of a restriction enzyme digested plasmid (5-20 pmol of 5'-ends, 0.1-0.5 units/pmol 5'-ends). Reduces re-ligation to < 0.5% compared to the untreated control.

One unit is the amount of the enzyme required to dephosphorylate 1 µg of linearized pUC57 DNA termini in 10 minutes at 37°C. The enzyme is purified to apparent homogeneity and is free of all contaminating endonucleases, exonucleases and ribonucleases

Unit Definition: One unit is the amount of the enzyme required to dephosphorylate 1 Â µg of linearized pUC57 DNA termini in 10 minutes at 37 °C.

 : Includes 10X reaction buffer: 200 mM Tris-HCl, pH 8,0, 100 mM MgCl₂; dilution buffer: 50 mM Tris-HCl, pH 8,0.