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Antarctic Phosphatase Reaction Buffer #B0289. Antarctic Phosphatase is active in all NEB restriction enzyme buffers only when supplemented with Antarctic Phosphatase Reaction Buffer, which provides Zn2+ required for enzyme activity The restriction enzyme should be heat inactivated at the same time as the phosphatase after digest and dephosphorylation Antarctic Phosphatase (AnP) is a heat labile alkaline phosphatase purified from a … Put the tube into the 37°C water bath for 1 hour. Antarctic Phosphatase activity is enhanced in the presence of monovalent salts. To the vector tube, add 5 μL of 10x Antarctic Phosphatase Buffer, 9μL of water, and 1 μL of Antarctic Phosphatase. Incubate the mixture at 37 °C for 1 h. Incubate the mixture at 70 °C for 10 min to inactivate Antarctic phosphatase. Purify DNA using a QIAquick PCR Purification Kit following the manufacturer's standard protocol. Concentration. AnP nonspecifically catalyzes the dephosphorylation of 5´ and 3´ ends of DNA and RNA phosphomonoesters. Antarctic Phosphatase Buffer, supplied by New England Biolabs, used in various techniques. Antarctic Phosphatase (AnP) is a heat-labile alkaline phosphatase purified from a recombinant source. Buffer supplied with Antarctic Phosphatase (NEB #M0289).
AnP nonspecifically catalyzes the dephosphorylation of 5´ and 3´ ends of DNA and RNA phosphomonoesters. 3. Catalog # Size. Rapid and irreversible heat inactivation eliminates unwanted activity Flexible reaction conditions (active in any restriction enzyme buffer, no clean-up required) Add 3 μl 10×PNK buffer, and 10 Units (1 μl) T4 polynucleotide kinase. A 1X concentration assures optimal enzyme activity. Also, AnP Hydrolyses ribo-, as well as deoxyribonucleoside triphosphates (NTPs … 10X Antarctic Phosphatase buffer; Procedure. Add 1μL Antarctic Phosphatase (probably should make final glycerol concentration less that 5%?) Storage Buffer: 25mM Tris-HCl (pH 7.5), 1mM MgCl2, 50% glycerol. Antarctic Phosphatase Protocol. Add Antarctic Phosphatase buffer to a final concentration of 1X to linearized vector sample. Antarctic Phosphatase (AnP) is a heat labile alkaline phosphatase purified from a recombinant source.
10X Antarctic Phosphatase Reaction Buffer RM20152 1X Antarctic Phosphatase Reaction Buffer Product Description Antarctic Phosphatase (AnP) is a heat labile alkaline phosphatase purified from a recombinant source. Add 5.5 μL of 10X Antarctic Phosphatase buffer and 2 μL (10 U) of Antarctic Phosphatase, mix the reaction, and return to 37°C for an additional 30 min. Add 1.5 μl 10x Antarctic phosphatase reaction buffer and 1 μl of Antarctic phosphatase. Incubate 60 mins at 37°C. A 1X concentration assures optimal enzyme activity.