Thermolabile UNG (Uracil N-Glycosylase)

For general laboratory use.

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 gel packs

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, 100 μg/ml BSA and 50 % [v/v] glycerol)

Concentration: 1 unit/μl

Description:
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 1 unit UNG completely digests 10⁴ to 10⁶ copies of U-containing DNA fragments in 2 min at 50°C.

Recommended assay:
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 complitely heat-inactivated at temperatures above 65°C in the following initial denataration step of the PCR.

Related products:

• qPCR Core Kits
• Dual labeled fluorescent probes
• Custom primers

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