Thermolabile UDG (500 U)

QIAGEN's Thermolabile Uracil-DNA Glycosylase is an enzyme designed for the removal of uracil from DNA and the prevention of carry-over contamination in PCR, qPCR, and RT-PCR workflows. The enzyme hydrolyzes the N-glycosylic bond between the deoxyribose sugar and the uracil base, leaving an apurinic or apyrimidinic (AP) site. It shows no activity on RNA substrates, making it suitable for use in RT-PCR reactions.

The protein is produced by a recombinant E. coli strain carrying the recombinant thermolabile Uracil DNA Glycosylase gene isolated from a marine bacterium, with a molecular weight of 26.2 KDa. The supplied concentration is 1,000 U/mL, and the storage temperature range is -25°C to -15°C. The enzyme is supplied in 50 mM Tris-HCl, 50 mM NaCl, 1 mM DTT, 0.1 mM EDTA, and 50% glycerol (pH 7.5 at 25°C), together with a 10X Reaction Buffer containing 700 mM Tris-HCl, 100 mM NaCl, 10 mM EDTA, and 1 mg/mL BSA (pH 8.0 at 25°C).

Quality specifications include a purity of >98%, a specific activity of 30,000 U/mg, single-stranded exonuclease activity of <1.0% released, double-stranded exonuclease activity of <1.0% released, no double-stranded endonuclease conversion, and E. coli DNA contamination of <10 copies. One unit is defined as the amount of enzyme required to release 1 nmol of uracil from dU-containing DNA in one hour at 37°C. The enzyme is completely inactivated by a 10-minute incubation at temperatures greater than 50°C in the 1x reaction buffer, with less than 0.5% activity remaining after such heat treatment. Typical usage involves adding 0.5 µL of the enzyme to a reaction containing uracil-containing DNA substrate (up to 0.1 µg) in a 20 µL total reaction volume, followed by incubation at 25–37°C for 30 minutes.