UNG (UDG) for preventing carry-over contaminations in RT-PCR assays
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
Concentration: 1 unit/μl
Extreme Thermolabile UNG (UDG) is used in one-step RT-PCR systems to prevent carry-over contamination with dU-containing DNA from previous reactions prior to the reverse transcription step. The enzyme is fully active at temperatures between 15 to 25°C and inactivated at 50°C.
Extreme Thermolabile UNG is a recombinant uracil-DNA glycosylase from marine bacterium BMTU 3346. UNG catalyzes the hydrolysis of the N-glycosidic bond between the uracil and sugar, leaving an apyrimidinic site in uracil-containing single or double-stranded DNA.
An amount of 1 unit UNG completely digests 104 to 106 copies of U-containing DNA fragments in 2 min at 50°C.
Use 0.2 units UNG per PCR assay (50 μl volume).
An UNG treatment at 20-25°C before starting the thermal cycling removes uracil residues from dU-containing DNA and prevents it from serving as template. Extreme Thermolabile UNG is completely heat-inactivated at temperatures above 50°C in the following reverse transcription step of the RT-PCR.
1 units/μl UNG in 20 mM Tris-HCl pH 9.0, 100 mM KCl, 0.1 mM EDTA, stabilizers, 50 % [v/v] Glycerol
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