Heat-labile Uracil-DNA Glycosylase (UDG) catalyzes the hydrolysis of the glycosidic bond of uracil, releasing free uracil and causing the degradation of DNA strands containing uracil bases. Compared to regular UDG, heat-labile UDG is more temperature-sensitive and easily inactivated by heating. This avoids the potential residual activity of regular UDG at room temperature, which could degrade dU-containing amplification products. The activity of heat-labile UDG on double-stranded DNA is lower than on single-stranded DNA. The enzyme is active on small dU-containing DNA oligonucleotides but not on RNA or normal dU-free DNA. Heat-labile UDG is active in the presence of Mg2+ or EDTA, as it does not require metal ions.
 
Product Numbers / Sizes: UDG-100µL (100 µL), UDG-200µL (200 µL), UDG-1mL (1000 µL)
Reaction Conditions:
Activation: Incubate for 30 minutes at a temperature of 25°C.
Inactivation: Incubate at a temperature of 50°C for 10 minutes.
Storage and Transport:
Storage Conditions: Store the product at a temperature of -20°C for up to two years.
Transportation: Transport the product at a temperature of 0°C or lower (≤0°C).
 
Enzyme Storage Buffer
20 mM Tris-HCl (pH 7.5)
100 mM KCl
0.1 mM EDTA
1 mM DTT
0.5% NP-40
0.5% Tween-20
50% glycerol.
 
Activity Definition: The activity of 1 unit (U) is defined as the amount of enzyme required to completely degrade 1 μg of dU-containing dsDNA within 30 minutes at 25°C.
 
Quality Control Exonuclease Activity Assay: Incubate 20 U of heat-labile UDG with 1 μg of Hind III-treated λDNA at 37°C for 10 hours. No change in DNA electrophoretic pattern should be observed.
 
Endonuclease Activity Assay: Incubate 20 U of heat-labile UDG with 1 μg of pUC19 plasmid DNA at 37°C for 1 hour. No change in DNA electrophoretic pattern should be observed.
 
Ribonuclease Activity Assay: Incubate 10 U of heat-labile UDG with 1 μg of 16S, 23S rRNA at 37°C for 1 hour. No change in RNA electrophoretic pattern should be observed.
Please see manual for more information.