Description:

The Quick Dephosphorylation Kit contains Quick CIP, a heat-labile recombinant version of calf intestinal alkaline phosphatase (CIP) purified from a recombinant source. Quick CIP nonspecifically catalyzes the dephosphorylation of 5´ and 3´ ends of DNA and RNA phosphomonoesters. It also hydrolyses ribo- and deoxyribonucleoside triphosphates (NTPs and dNTPs). Quick CIP is useful in many applications that require the dephosphorylation of DNA or RNA ends. In cloning, dephosphorylation prevents re-ligation of linearized plasmid DNA. The enzyme can quickly dephosphorylate 5´ protruding, 5´ recessed, and blunt ends in just 10 minutes. Quick CIP may also be used to degrade unincorporated dNTPs in PCR reactions to prepare templates for DNA sequencing or SNP analysis. Quick CIP is completely and irreversibly inactivated by heating it at 80°C for 2 minutes, unlike wild type CIP, which is heat-inactivatable. This makes removal of Quick CIP prior to ligation or end-labeling unnecessary.1X CutSmart Buffer:50 mM Potassium Acetate20 mM Tris-acetate10 mM Magnesium Acetate100 µg/ml BSApH 7.9 @ 25°C

Kit Components

The following reagents are supplied with this product:

	Store at (°C)	Concentration
CutSmart® Buffer	-20	10X
Quick CIP	-20

Notes:

Quick CIP, as are most alkaline phosphatases, is a Zn2+ and Mg2+-dependent enzyme. Our formulation has tightly bound zinc atoms in the active center and does not require supplemental zinc or other additives.Quick CIP is also active in 1X NEBuffers 1.1, 2.1, 3.1 as well as NEBuffers 1, 2, 3 and 4.Quick CIP activity is enhanced in the presence of monovalent salts.Quick CIP is inhibited by metal chelators (e.g. EDTA), inorganic phosphate and phosphate analogs.The Quick CIP activity is decreased in the presence of reducing agents (DTT, β-mercaptoethanol).The source for Quick CIP is a Pichia pastoris clone that carries the calf intestinal alkaline phosphatase gene from calf intestinal mucosa.The storage conditions for Quick CIP are 25 mM Tris-HCl, 1 mM MgCl2, 0.1 mM ZnCl2, 50% Glycerol, pH 7.5 @ 25°C.Quick CIP can be heat inactivated for 2 minutes at 80°C.Quick CIP is a homodimer. The molecular weight of the monomer is 56 kDa.