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Product description: Uracil-DNA Glycosylase (UNG),E.coli is a recombinant protein in E.coli. The enzyme hydrolyzes uracil-glycosidic bonds at U-DNA in single- or double-strand DNA excising uracil and creating alkali sensitive abasic sites in the DNA. The enzyme is inactive on RNA and native, uracil-free DNA. Since Uracil-DNA Glycosylase (UNG), heat-labile has no metal ion requirements, it is fully active in the presence of EDTA.
Source: Recombinant E.coli
Concentration and Size: 1U/μL
Unit Definition: One unit is defined as the amount of enzyme required to catalyze the release of 1 nmol uracil from uracil-containing DNA in 1h at 37℃.
Storage Buffer: 20mM Tris-HCl (pH8.0), 0.1mM EDTA, 100mM KCl, 1mM DTT, 0.5%(v/v) Tween-20, 0.5%(v/v) NP-40, 50%(v/v) glycerol.
1. UDG, E.coli is fully active at 37℃, 42℃ and 50℃.
2. Uracil-DNA Glycosylase from E.coli has to be inactivated for 10min at 95℃.
3. It is suggested that adding 0.1U～0.5U Uracil-DNA Glycosylase (UNG), E.coli to 50μL PCR System.
4. Incubate at 37℃ for 15~30min to release uracil from the DNA.
Storage Conditions: -20℃
Quality Assurance: Free of Endonuclease, Exonuclease, Nickase activities and RNase free
PCR carry-over prevention: 5ng dUTP-containing PCR products as template to amplify 200bp target DNA.
Before the amplification reaction, add 0.1 unit of Uracil-DNA Glycosylase (UNG), E.coli to PCR mixes, after 10min at 37℃, no reamplification is possible.
Physical Purity: ＞95% by SDS-PAGE.
1. Lindahl T, Ljungquist S, Siegert W, et al. The Journal of Biological Chemistry. 252, 3286-94(1977).
2. Longo MC，Berninger MS，Hartley JL. Gene. 93, 125-8(1990).