Alkaline Phosphatase, Calf Intestinal (HC) (1,000u)

Alkaline Phosphatase, Calf Intestinal (CIAP), catalyzes the hydrolysis of 5´-phosphate groups from DNA, RNA, and ribo- and deoxyribonucleoside triphosphates. This enzyme is used to prevent recircularization and religation of linearized cloning vector DNA by removing phosphate groups from both 5´-termini and may also be used for the dephosphorylation of 5´ phosphorylated ends of DNA or RNA for subsequent labeling with [32P]ATP and T4 Polynucleotide Kinase. CIAP is active on 5´ overhangs, 5´ recessed and blunt ends.

Features - Benefits

• Available at High Concentration: Cat.# M2825 contains 1,000 units of CIAP at 20u/μl.
• Blue/White Cloning Qualified: Promega's blue/white cloning assay provides a higher level of quality control for enzymes used in cloning applications.
• Provided with 10X Reaction Buffer: 0.5M Tris-HCl (pH 9.3 at 25°C), 10mM MgCl₂, 1mM ZnCl₂, 10mM spermidine.

Applications

• Preventing religation of linearized cloning vehicle DNA by removing phosphate groups from both 5´ termini.
• Removing 5´ phosphate groups prior to end-labeling with T4 Polynucleotide Kinase.

For more information, see the Protocols & Applications Guide.

References

1. Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
2. Seeburg, P.H. et al. (1977) Nature 270, 486–94.
3. Ullrich, A. et al. (1977) Science 196, 1313–9.
4. Meyerowitz, E.M. et al. (1980) Gene 11, 271–82.
5. Grosveld, F.G. et al. (1981) Gene 13, 227–37.