Antarctic Thermolabile UDG (Uracil-DNA Glycosylase) catalyzes the release of freeuracil from uracil-containing single-stranded ordouble-stranded DNA. The resulting abasic sitesare susceptible to hydrolytic cleavage at elevatedtemperature and high pH. This enzyme is sensitiveto heat and can be rapidly and completely inactivatedat temperatures above 50°C.Figure 1:Evaluation of RT-qPCR carryover preventionTo evaluate the capacity of carryover product digestion, a uracil-containing RT-qPCR product was generated using the Luna One-Step Probe RT-qPCR kit. Approximately 105 copies of the uracil-containing product were used as template for subsequent qPCR reactions using three different RT-qPCR master mixes (all containing UDG). Following manufacturer’s recommended protocols, incubation at 25˚C at the start of the 2nd reaction enables UDG to degrade the dU-containing input, reducing its ability to contribute to a (false) positive signal. The ΔCq value is the cycle difference between carryover treatment and no carryover treatment of the same input. Larger ΔCq values indicate more efficient carryover product digestion. After decontamination using the Luna mix, full product digestion was observed in one sample and a large ΔCq was observed in the second. Note that the amount of DNA present in true contamination events is difficult to assess/predict.
Product Source
An E. coli strain that carries the cloned UDG gene from a psychrophilic marine bacterium
This product is related to the following categories:
DNA Repair Enzymes and Structure-specific Endonucleases Products
This product can be used in the following applications:
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