E.coli uracil-DNA glycosylase (UDG) catalyzes the release of free uracil from uracil-containing DNA. UDG efficiently hydrolyzes uracil from single-stranded or double-stranded DNA, but not from oligomers (6 or fewer bases)
Applications:
- site-directed mutagenesis;
- as a probe for protein-DNA interaction studies;
Treatment of 1 ugof uracil-containing DNA with 1 unit of UDG for 10 minutes at 37oC renders the DNA incapable of being copied by DNA polymerase. The enzyme can be 95% heat killed by incubation at 95oC for 10 minutes.
Unit
One unit is defined as the amount of enzyme that catalyzes the release of 60 pmol of uracil per minute from double-stranded, uracil-containing DNA. Activity is measured by release of [3H]-uracil in a 50 ul reaction containing 0,2 ug DNA (104 -105 cpm/ug) in 30 minutes at 37oC.
Reaction buffer
SE-buffer Uracil DNA Glycosylase, (20 mM Tris-HCl (pH 8.0 at 25°C); 1 mM EDTA; 1 mM DTT)
Optimal temperature
37oC
Storage conditions
10 mM Tris-HCl (pH 7.6); 50 mM NaCl; 1 mM EDTA; 1 mM DTT; 50% glycerol. Store at -20oC.
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