Thermostable dUTPase
Available from http://www.labsupplymall.com
Thermostable dUTPase (Cloned)
Thermostable dUTPase (pyrococcus fruriosus) was prepared and purified from Escherichia coli expression system. dUTPase is specific for dUTP and is critical for the fidelity of DNA replication and repair. dUTPase hydrolyzes dUTP to dUMP and pyrophosphate, simultaneously reducing dUTP levels and providing the dUMP for dTTP biosynthesis.
Thermostable dUTPase used for PCR with Pfu DNA polymerase improves the efficiency and longer targets of PCR reaction.
Unit Definition
One unit of enzyme catalyzes hadrylazation of 10 nanomoles of dUTP to dUMP in one hour at 85 Centigrade
Activity
(A) Measured by its ability to hydrolyze dUTP to dUMP in reaction buffer 20mM Hepers pH7.5, 150mM KCI, 5mM MgCl2, 10mM dUTP at 85 Centigrade for 1 hour. The PPi production was quantified by using the enzymatic determination kit from Sigma.
(B) Enhancing PCR amplication: 50ul of Pfu PCR reaction system with 1-3u of dUTPase to amplify genomic DNA target up to 15-19 kb in length.
Specific activity: 103 u/ug
Formulation: dUTPase is supplied in 20mM Tris-HC1 (pH 8.2), 1mM DTT, 0.1mM EDTA, 100mM KC1, 0.1% Nonidet P40, 0.1% Tween 20 and 50% glycerol at concentration of 1u/ul of the enzyme.
Reference
1. Dabrowski S. and Ahring B.K. protein Exp. Pur. 2003,31,72-78
2. Hogrefe H. et al. Proc. Natl. Acad. Sci. USA 2002,99,596-601
