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Products » Molecular Biology Products » DNA/RNA Modifying Enzymes » Enzyme : Uracil DNA Glycosylase
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Description:E. coli Uracil DNA Glycosylase (UNG) catalyses the release of free uracil from uracil-containing DNA. UNG efficiently hydrolyzes uracil from single-stranded or double-stranded DNA, but not from oligomers (6 or fewer bases).
Source:An E. coli strain that carries the UNG gene from E. coli.
Applications:
- Glycosylase mediated single nucleotide polymorphism detection (GMPD)
- Site-directed mutagenesis
- As a probe for protein-DNA interaction studies
- Rapid and efficientcloning of PCR products
- Elimination carry-over contamination in PCR
Quality Control: Activity, SDS-PAGE (purity), 16-hour incubation, exonuclease and endonuclease activity
Unit Definition:One unit is defined as the amount of enzyme that catalyzes the release of 60 pmol of uracil per minute from double-stranded, uracil-containing DNA in a total reaction volume of 50 μl in 30 minutes at 37°C in 1X Uracil DNA Glycosylase Reaction Buffer with 1 unit or uracil DNA Glycosylase and 0.2 μg [3H]-uracil DNA (104-105 cpm/μg).
10X UNG Reaction Buffer:200 mM Tris-HCl (pH8.0 at 25°C), 10 mM Dithiothreitol, 10 mM EDTA.
Reaction Conditions:1X UNG Reaction Buffer, incubate at 37°C.
Inhibition and Inactivation:Inactivated by heating at 95°C for 10 min. Enzyme activity is partially restored at temperatures lower than 55°C.
Storage Buffer: UNG in 10 mM Tris-HCl (pH7.4 at 25°C), 50 mM KCl, 1 mM Dithiothreitol, 0.1 mM EDTA, 0.1 mg/ml BSA, 50% Glycerol
Concentration: 5 U/µl
Storage:Store at -20°C.
Note:UNG is active over a broad pH range with an optimum at pH 8.0, does not require divalent cation, and is inhibited by high ionic strength (>200 mM).
The abasic sites formed in DNA by UNG may be cleaved by heat, alkali-treatment or endonucleases that cleave specifically at abasic sites.
MSDS: 20080228032005 (PDF)