Add 1μL Antarctic Phosphatase (probably should make final glycerol concentration less that 5%?) Add 1.5 μl 10x Antarctic phosphatase reaction buffer and 1 μl of Antarctic phosphatase. Antarctic Phosphatase can also be used in NEBuffers 1, 2, 3 or 4 as well as the unique NEBuffers for EcoR I and BamH I ONLY when supplemented with 10X Antarctic Phosphatase Reaction Buffer to a final concentration of 1X.
After, place it in the 82°C heating block for 20 minutes. The change in absorbance at 405 nm is monitored (3050 µL reaction volume). New England Biolabs supplies a 10X reaction buffer with all of its enzymes. Bioz Stars score: 99/100, based on 13 PubMed citations. 10X Antarctic Phosphatase buffer; Procedure. A 1X concentration assures optimal enzyme activity. Incubate 60 mins at 37°C. Order:email@example.com Forresearchpurposesonly.Notfortherapeuticordiagnosticpurposesinhumansoranimals. AnP nonspecifically catalyzes the dephosphorylation of 5´ and 3´ ends of DNA and RNA phosphomonoesters. Antarctic Phosphatase (AnP) is a heat-labile alkaline phosphatase purified from a recombinant source. A 1X concentration assures optimal enzyme activity. Antarctic Phosphatase Reaction Buffer #B0289. 3 μl [γ-32 P]-ATP. 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 Purify DNA using a QIAquick PCR Purification Kit following the manufacturer's standard protocol. To the vector tube, add 5 μL of 10x Antarctic Phosphatase Buffer, 9μL of water, and 1 μL of Antarctic Phosphatase. Catalog # Size. Add Antarctic Phosphatase buffer to a final concentration of 1X to linearized vector sample.
Concentration. We do this to prevent the vector from closing up again without any insert. Buffer supplied with Antarctic Phosphatase (NEB #M0289). Incubate the mixture at 37 °C for 1 h. Incubate the mixture at 70 °C for 10 min to inactivate Antarctic phosphatase.
Buffer supplied with Antarctic Phosphatase (NEB #M0289).
3. 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. 4. At a 1X concentration this reaction buffer assures optimal activity of the enzyme. Antarctic Phosphatase is also active in all restriction enzyme NEBuffers 1.1, 2.1, 3.1 and CutSmart ® Buffer only when supplemented with 1/10 volume of the 10X Antarctic Phosphatase Reaction Buffer. Antarctic Phosphatase activity is enhanced in the presence of monovalent salts. Antarctic Phosphatase (AnP) is a heat labile alkaline phosphatase purified from a … ZERO BIAS - … Assay Conditions: The reaction mixture contains 100mM glycine, pH 10.4, 1mM MgCl 2, 1mM ZnCl 2, 10mM p-nitrophenyl phosphate, and 0.001-0.1 units of Shrimp Alkaline Phosphatase (SAP).
Put the tube into the 37°C water bath for 1 hour. Antarctic Phosphatase Buffer, supplied by New England Biolabs, used in various techniques. Rapid and irreversible heat inactivation eliminates unwanted activity Flexible reaction conditions (active in any restriction enzyme buffer, no clean-up required) 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 …
Antarctic Phosphatase Protocol. Add 3 μl 10×PNK buffer, and 10 Units (1 μl) T4 polynucleotide kinase. 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. Storage Buffer: 25mM Tris-HCl (pH 7.5), 1mM MgCl2, 50% glycerol.