Prevention of carry-over contaminations
For in vitro use only!
Unit Definition: One unit of enzyme catalyzes the degradation of 1 μg single-stranded uracil-containing DNA at 37 °C in 60 min.
Shipping: shipped on blue ice
Storage Conditions: store at -20 °C
avoid freeze/thaw cycles
Shelf Life: 12 months
Form: liquid (Supplied in 20 mM Tris-HCl pH 8.0, 50 mM NaCl,1 mM EDTA, 1 mM DTT, 50 μg/ml BSA and 50 % [v/v] glycerol)
Concentration: 1 unit/μl
Thermolabile UNG is used in real-time PCR to prevent carry-over contamination of dU-containing DNA from previous reactions. Uracyl N-Glycosylase (UNG, UDG) catalyses the release of uracil from single and double stranded uracyl-containing DNA. The resulting abasic sites are susceptible to hydrolytic cleavage at elevated temperatures.
An amount of 0.1 units UNG can completely destroy up to 200 ng dU-containing DNA in 2 min at 50°C.
Add 0.2 μl (0.2 units) UNG for each 50 μl of master mix and vortex thoroughly. The preparation of a master mix is crucial in quantitative PCR reactions to reduce pipetting errors.
An UNG treatment of 2 min at 50°C at the onset of thermal cycling removes uracil residues from dU-containing DNA and prevents it from serving as template. UNG is easily heat-inactivated at temperatures above 65°C in the following initial denataration step of the PCR.