Calf-intestinal alkaline phosphatase
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Calf Intestinal Alkaline Phosphatase (CIAP/CIP) is an enzyme that catalyzes the 5' phosphate group's removal, Dephosphorylation, from DNA [1] [2]. This enzyme is frequently used in DNA sub-cloning, because DNA fragments that lack the 5' phosphoryl termini cannot self-ligate [3]. This is used to prevent recircularization and religation of linearized cloning vehicle DNA by removing phosphate groups from both 5´-termini. Preventing self-ligation is important both in improving the yield of properly ligated product and in reducing the background of improperly self-ligated contaminant.
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Notes
- ^ Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
- ^ Seeburg, P.H. et al. (1977) Nucleotide sequence and amplification in bacteria of structural gene for rat growth hormone. Nature 270, 486–94
- ^ Ullrich, A. et al. (1977) Rat insulin genes: Construction of plamids containing the coding sequences. Science 196, 1313–9.
References
- Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
- Seeburg, P.H. et al. (1977) Nucleotide sequence and amplification in bacteria of structural gene for rat growth hormone. Nature 270, 486–94
- Ullrich, A. et al. (1977) Rat insulin genes: Construction of plamids containing the coding sequences. Science 196, 1313–9.