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Uracil-DNA Glycosylase*
* Use of this enzyme in certain applications may be covered by patents and may require a license.

#EN0361 Supplied with: 10X Reaction Buffer	200 u (1 u/µl) 0.4 ml
#EN0362 Supplied with: 10X Reaction Buffer	5x200 u (1 u/µl) 5x0.4 ml
Related Documents (in pdf, ~40-60 KB): Certificate of Analysis: #EN0361, #EN0362 MSDS (English) MSDS (English-USA) MSDS (German)

 Description
The Uracil-DNA Glycosylase catalyzes the hydrolysis of the N-glycosylic bond between the uracil and sugar, leaving an apyrimidinic site in uracil-containing single or double-stranded DNA. Shows no activity on RNA (1).

Source
E.coli K12 cells.

Molecular Weight
25.6 kDa monomer.

Applications

• Glycosylase mediated single nucleotide polymorphism detection (GMPD) (2).
• Site-directed mutagenesis (3).
• As a probe for protein-DNA interaction studies (4).
• SNP genotyping.
• Rapid and efficient cloning of PCR products (5).
• Elimination carry-over contamination in PCR (6).

Quality Control
The absence of endodeoxyribonucleases, exodeoxyribonucleases, phosphatases and ribonucleases confirmed by appropriate quality tests.

Concentration
1 u/µl

Definition of Activity Unit
One unit of the enzyme catalyzes the release 1 nanomole of uracil from uracil-containing DNA template in 60 min at 37°C.

Storage Buffer
The enzyme is supplied in: 30 mM HEPES-KOH (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.05% (v/v) Tween 20 and 50% (v/v) glycerol.

10X Reaction Buffer
200 mM Tris-HCl (pH 8.2 at 25°C), 10 mM EDTA, 100 mM NaCl.

Inhibition and Inactivation
Inactivated by heating at 95°C for 10 min. Enzyme activity is partially restored at temperatures lower than 55°C.

Note
The abasic sites formed in DNA by Uracil-DNA Glycosylase may be subsequently cleaved by heat, alkali-treatment or endonucleases that cleave specifically at abasic sites.

Related Products

Exonuclease III Endonuclease IV, E.coli dUTP Water, nuclease-free

References

1. Lindahl, T., et al., DNA N-Glycosidases, J. Biol. Chem., 252, 10, 3286-3294, 1977.
2. Vanghan, P., McCarthy, T.V., A novel process for mutation detection using uracil DNA glycosylase, Nucleic Acids Res., 26, 810-815, 1998.
3. Kunkel, T.A., Rapid and efficient site-specific mutagenesis without phenotypic selection, Proc. Natl. Acad. Sci. USA, 82, 488-492, 1985.
4. Devchand, P.R., et al., Uracil-DNA glycosylase as a probe for protein-DNA interactions, Nucleic Acids Res, 21, 3437-3443, 1993.
5. Booth, P.M., et al., Assembly and cloning of coding sequences for neurotrophic factors directly from genomic DNA using polymerase chain reaction and uracil DNA glycosylase, Gene, 146, 303-308, 1994.
6. Longo, M.C., et al., Use of uracil DNA glycosylase to control carry-over contamination in polymerase chain reactions, Gene, 93, 125-128, 1990.