Uracil-DNA Glycosylase (UNG), Cod is a thermolabile recombinant enzyme produced in E. coli (ung-) by a modified ung gene derived from Atlantic Cod. It degrades uracil-containing single- and double-stranded DNA, but not RNA or thymidine-containing DNA, by hydrolyzing the N-glycosidic bond between deoxyribose sugar and the base in uracil. This generates alkaline-sensitive apyramidinic sites in the DNA that will be cleaved upon a combination of alkaline conditions and high temperature.
Pretreatment of samples with UNG prevents PCR carryover contamination in labs that substitute dUTP in place of dTTP during all amplification reactions. PCR products containing uracil become substrates for UNG and will be degraded if they are present in subsequent reaction mixtures subjected to UNG treatment. Only DNA templates containing thymidine are not degraded by the treatment and will be amplified.
Recombinant cod UNG is irreversibly heat inactivated, which enables long-term storage and subsequent analysis of post-PCR amplicons in applications such as cloning and sequencing. AMRESCO’s UNG is compatible with PCR, qPCR and one-step qRT-PCR and works in all commercially available master mixes. All amplification reactions must use dUTP containing dNTP mixtures in order for the UNG decontamination method to be effective.
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