Activity, DNase, RNase, endonuclease, end-labeling, blue/white cloning assay.
Recombinant E. coli strain.
20mM Tris-HCl (pH 7.5), 25mM KCl, 2mM DTT, 0.1mM EDTA, 0.1μM ATP and 50% (v/v) glycerol.
Store at –20°C.
One unit is defined as the amount of enzyme required to catalyze the transfer of 1nmol of phosphate to the 5´-OH end of a polynucleotide from [γ-32P]ATP in 30 minutes at 37°C. The reaction conditions are: 40mM Tris-HCl (pH 7.5 at 25°C), 10mM MgCl2, 5mM DTT, 0.1mM [γ-32P] ATP and 0.5mM 5´-OH polynucleotide end concentration.
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