New England Biolabs, M0281L, Uracil Glycosylase Inhibitor (UGI)

Related Categories DNA Repair Enzymes and Structure-specific Endonucleases Specification Unit Definition One unit of UGI is defined as the amount of protein required to inhibit one unit of E. coli UDG in 1 hour at 37°C in a total reaction volume of 50 μl. One unit of UDG is the amount of enzyme which will catalyze the release of 60 pmol of uracil per minute from double stranded, uracil-containing DNA. Reaction Conditions 1X UDG Reaction Buffer Incubate at 37°C 1X UDG Reaction Buffer 20 mM Tris-HCl 1 mM DTT 1 mM EDTA (pH 8 @ 25°C) Storage Buffer 10 mM Tris-HCl 50 mM KCl 1 mM DTT 0.1 mM EDTA 200 µg/ml BSA 50% Glycerol pH 7.4 @ 25°C Heat Inactivation 70°C for 10 minutes FAQ Q: What is the nature of the Uracil Glycosylase Inhibitor? A: UGI is a protein. Q: How does UGI inhibit UDG? A: UGI is capable of dissociating UDG-DNA complexes. It binds with a 1:1 stoichiometry to UDG. Q: Does UGI inhibit the activity of all UDGs? A: UGI is able to inhibit the activity of E coli UDG, as well as the activity of UDGs from other species. However, UGI is not capable of inhibiting the activity of Afu UDG. Q: What is the molecular weight of UGI? A: UGI has a molecular weight of 9.5 kDa. Q: Why is it necessary to use UGI in a reaction? A: UDG remains partially active following heat treatment at 95°C. UGI can be added to the reaction to prevent subsequent degradation of the product DNA.