Klenow fragment is a DNA-dependent polymerase with 3'-5' exonuclease activity. It lacks 5'-3' exonuclease activity of the native enzyme and is suited for random-primed labelling of DNA with random oligonucleotides and for filling-in of 5' protruding ends to blunt-ends.
One unit is the amount of enzyme that incorporates 10 nmoles of deoxynucleotides into acid-insoluble form in 30 minutes at 37░C.
20 mM Tris-HCl pH 7.5, 100 mM NaCl, 0.1 mM EDTA, 2 mM DTT, 0.1 mg/ml BSA; 50% glycerol (v/v)
Store Klenow Enzyme below 0║C, preferably at -20║C, in a constant temperature freezer.
10X REACTION BUFER
100 mM Tris-HCl pH 7.5, 100 mM MgCl2 , 10 mM DTE
(= buffer L for restriction endonucleases!)
T4 DNA Ligase, dNTP