- Phosphate group removal to reduce self-ligation and vector background.
- Dephosphorylation of DNA 5' termini DNA prior to kinase labeling reactions
E. coli strain C75
Bacterial Alkaline Phosphatase (E.coli
) is extremely stable and is not completely heat-inactivated; therefore, to inactivate the enzyme, reaction products need to be purified twice via phenol-chloroform extraction followed by ethanol precipitation.
Supplied with 10X Reaction Buffer [500 mM Tris-HCl (pH 9.0), 10 mM MgCl2]
- Reid, T.W. & Wilson, I.B. (1971) The Enzymes 3:373-416.
- Takanami, M. (1967) J. Mol. Biol. 23:135-148.
One unit is defined as the amount of enzyme required to generate 1 µmol/min of p-nitrophenol from p-nitrophenylphosphate at 25°C and pH 8.0.