Properties & Usage
Unit Definition
One unit is defined as the amount of enzyme required to cleave 20 pmol of a 34 mer oligonucleotide duplex containing a single AP site* in a total reaction volume of 10 µl in 1 hour at 37°C.
*An AP site is created by treating 20 pmol of a 34 mer oligonucleotide duplex containing a single uracil residue with 1 unit of Uracil-DNA Glycosylase (UDG) for 2 minutes at 37°C. Reaction Conditions
1X NEBuffer™ 4
Incubate at 37°C
1X NEBuffer™ 4
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
1 mM DTT
(pH 7.9 @ 25°C)
Storage Buffer
10 mM Tris-HCl
50 mM NaCl
1 mM DTT
0.05 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 8 @ 25°C
Heat Inactivation
65°C for 20 minutes Unit Assay Conditions
1X NEBuffer 4 containing 20 pmol of fluorescently labled oligonucleotide duplex in a total reaction volume of 10 μl.