NZY Thermolabile Uracil-DNA Glycosylase (UDG)

NZY Thermolabile Uracil-DNA Glycosylase is an engineered version of an Uracil-DNA Glycosylase (UDG) from a cold-adapted organism that, at low temperatures (20-25°C), catalyses the release of free uracil from uracil-containing DNA templates. The enzyme efficiently hydrolyses uracil from single-stranded or doublestranded DNA (although it is more active on ssDNA), but not from RNA. The enzyme excises uracil, thus creating alkali-sensitive abasic sites in the DNA. These abasic sites can be hydrolysed by heat and thus the contaminant dU-containing LAMP/PCR products from previous experiments can no longer serve as amplification templates. Since the enzyme is inactivated at temperatures of 45°C or higher, NZY Thermolabile Uracil-DNA Glycosylase is useful in a variety of applications where carry-over contamination can be a problem. These include PCR/qPCR or LAMP (since the LAMP reaction takes place at 65°C), as well as these reactions merged with reverse transcription (RT) RT-LAMP or RT-PCR/qPCR (given that RT usually takes place at 45-55°C). At 45°C, NZY Thermolabile Uracil-DNA Glycosylase is completely inactivated and thus will not target the newly generated uracil-containing cDNA/DNA chains in RT or LAMP/PCR reactions, respectively. An incubation step with NZY Thermolabile Uracil-DNA Glycosylase for 2-5 min at 25°C before PCR/qPCR reactions is all that is required to prevent the amplification of contaminant LAMP/PCR products carried over.