"Glycosylase" Related Products

Uracil DNA Glycosylase(UNG)

Cat. No.: CMC13
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 efficient cloning of PCR products; Elimination carry-over contamination in PCR.
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℃ in 1× Uracil DNA Glycosylase Reaction Buffer with 1 unit of uracil DNA Glycosylase and 0.2 μg [3H]-uracil DNA (104-105 cpm/μg).
Quality Control: Activity, SDS-PAGE (purity), 16-Hour Incubation, Exonuclease and Endonuclease Activity.
Storage: UNG in 10 mM Tris-HCl (pH 7.4 at 25°C), 50 mM KCl, 1 mM Dithiothreitol, 0.1 mM EDTA, 0.1 mg/ml BSA, 50% Glycerol should be stored at -20℃.
10× UNG Reaction Buffer: 200mM Tris-HCl (pH 8.0 at 25°C), 10mM Dithiothreitol, 10mM EDTA.
Reaction Conditions: 1× UNG Reaction Buffer, incubate at 37℃.
Inhibition and Inactivation: Inactivated by heating at 95℃ for 10min. Enzyme activity is partially restored at temperatures lower than 55°C.
Usage notes: 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.

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