Synonyms Uracil DNA Glycosilase, Uracil DNA Glycosylase, UNG.
Description E.Coli Uracil DNA Glycosilase (UNG) catalyses the release of free Uracil from Uracil-containing DNA. UNG efficiently hydrolyzes uracil from signle-stranded or double-stranded DNA, but not from oligomers (6 fewer bases).
Source Escherichia Coli strain that carries the UNG gene from E.coli.
Physical Appearance Sterile Filtered colorless solution.
Formulation UNG solution in 10mM Tris-Hcl (pH-7.4 at 25℃), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.1 mg/ml BSA and 50% glycerol.
Stability Uracil DNA Glycosilase although stable at 15℃ for 1 week, should be stored desiccated below -18℃.
Please prevent freeze-thaw cycles.
Applications Glycosilase 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.
Specific Activity The Specific Activity was found to be 5U/µl.
Unit Definition 1 Unit of the enzyme catalyzes the release of 1 nanomole of uracil-containing DNA template in 60 min at 37℃.
10×UNG Reaction Buffer 200mM Tris-HCl (pH 8.0 at 25℃), 10mM DTT and 10mM EDTA.
Reaction Conditions 1X UNG Reaction Buffer, incubate at 37℃.
UNG is active over a broad pH rabge with an optimum at pH-8.0, doesn't require divalent cation, and is inhibited by high ionic strength (>200mM). The abasic sites formed in DNA by UNG may be cleaved by heat, alkali-treatment or endonucleases that cleave specifically at abasic sites.
Inhibition and Inactivation Inactivated by heating at 95℃ for 10min. Enzye activity is partially restored at temperatures lower than 55℃.
Usage CHI's products are furnished for LABORATORY RESEARCH USE ONLY. They may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.