Uracil-DNA Glycosylase (UNG), E.coli

Product Information

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: 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.

Key points of operation

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.

Security Information

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.

Reference

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).