Cryophile Uracil DNA Glycosylase
Cryophile Uracil DNA Glycosylase or Uracil-N glycosylase (UNG) hydrolyses the N-glycosylic bond between the deoxyribose sugar and the base in uracil- containing DNA leaving a basic (apyrimidinic) site in DNA. Cryophile Uracil DNA Glycosylase is not active on uracil in RNA. The high activity combined with a fast irreversible heat-inactivation step makes the enzyme ideal for contamination control in PCR and RT-PCR reactions. Unlike all other available UNG enzymes, Cryophile Uracil DNA Glycosylase does not reactivate after heat-inactivation. This enables contamination control in amplification reactions also where subsequent post-PCR analysis is required.
- The only UNG that can be completely and irreversibly heat inactivated
- Heat-labile without any addition of agents or inhibitors
- Eliminates carry-over contamination in both PCR and RT-PCR
- Enables downstream post-PCR analysis such as cloning and sequencing
Arctic cod origin, recombinantly produced in E. coli.
Cryophile Uracil DNA Glycosylase is completely and irreversibly inactivated by incubating the enzyme at 55°C for 20 min.
Minimum shelf life is 2 years at -20°C. Storage at 4°C is possible for at least 6 months. The enzyme also tolerates multiple freeze-thaw cycles.
Cryophile Uracil DNA Glycosylase highly pure and is tested free of contaminating nucleases.
> 500 000 Units/mg
One unit of UNG is defined as the amount of enzyme required to release 1 nmol uracil from uracil-containing DNA per hour at 37°C
Use of Cryophile Uracil DNA Glycosylase in RT-PCR
A prerequisite for using UNG in RT-PCR contamination control, is that the enzyme is sufficiently heat-labile to quickly inactivate at the temperatures used for reverse transcription. The easily heat-inactivatable Cryophile Uracil DNA Glycosylase makes it possible to use contamination control also in RT-PCR, being able to remove more than 108 copies of contaminating DNA without affecting the sensitivity (Cq) of the assay. Applying Cryophile Uracil DNA Glycosylase in an RT-PCR reaction is done by a 5 min Cryophile Uracil DNA Glycosylase preincubation step at room temperature prior to the amplification reaction. Cryophile Uracil DNA Glycosylase does not affect sensitivity of qRT-PCR.
PCR product stability after Cryophile Uracil DNA Glycosylase treatment
The complete and irreversible heat-inactivation of Cryophile Uracil DNA Glycosylase enables the combination of contamination control and an array of post-PCR applications such as cloning, sequencing or genotyping. As Cryophile Uracil DNA Glycosylase is the only commercially available UNG that does not reactivate post-amplification, PCR products remain intact and can be stored for a prolonged period of time before further analysis.
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