Purified from calf intestinal mucosa, this enzyme catalyses the hydrolysis of phosphate monoesters. Calf intestine alkaline phosphatase, rather than bacterial enzyme is often chosen due to its high turnover rate and milder conditions of inactivation. AP can be used to remove 5'-phosphates from DNA or RNA prior to 5'-end labelling. AP is also used to remove 5'- phosphates from linearised vector molecules to prevent self-ligation of the vector during cloning procedures. The enzyme is tested for endonuclease and ribonuclease contamination.
One unit is the amount of enzyme required to hydrolyse 1nm of p-nitrophenylphosphate in 1 min at 37°C in a buffer of 1.0M diethanolamine, 10mM p-nitrophenylphosphate, 0.25mM MgCl2 , pH 9.8.
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