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Uracil-DNA Glycosylase

B buffer for 100% activity G buffer for 100% activity LO certified Tango buffer for 100% activity Thermal inactivation at 95°C in 10 min

Uracil-DNA Glycosylase (UDG, UNG) catalyzes the hydrolysis of the N-glycosylic bond between uracil and sugar in DNA and prevents carry-over of DNA in PCR reactions.
  
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Thermo Scientific Uracil-DNA Glycosylase (UDG, UNG) catalyzes the hydrolysis of the N-glycosylic bond between uracil and sugar, leaving an apyrimidinic site in uracil-containing single or double-stranded DNA (see Figure 1 in Supporting Data). The enzyme shows no activity on RNA (see Reference 1).

Highlights

  • Active in Thermo Scientific buffers for restriction enzymes and thermophilic polymerases

Applications

  • Control of carry-over contamination in PCR (see​ Reference 2)
  • Glycosylase mediated single nucleotide polymorphism detection (GMPD) (see​ Reference 3)
  • Site-directed mutagenesis (see​ Reference 4)
  • As a probe for protein-DNA interaction studies (see​ Reference 5)
  • SNP genotyping
  • Cloning of PCR products (see​ Reference 6)
  • Generation of single strand overhangs of PCR products and cDNA

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. UDG (UNG) is active in the presence or absence of divalent cations.

Use of this enzyme in certain applications may be covered by patents and may require a license.

  
Storage Condition-20 C
HazardousNo
10X Reaction Buffer200 mM Tris-HCl (pH 8.2 at 25°C), 10 mM EDTA, 100 mM NaCl
Concentration1 U/µL
Definition of Activity UnitOne unit of the enzyme catalyzes the release 1 nm of uracil from uracil-containing DNA template in 60 min at 37°C.
InactivationInactivated by heating at 95°C for 10 min. Enzyme activity is partially restored at temperatures lower than 55°C. Therefore, put PCR products on ice after PCR and load directly on a gel.
InhibitionInhibitors: Ugi protein from the Bacillus subtilis phage PBS2, protein p56 from the Bacillus subtilis phage phi29 (see Reference 7).
Molecular Weight25.6 kDa monomer
Quality ControlThe absence of endo-, exodeoxyribonucleases, phosphatases, and ribonucleases confirmed by appropriate quality tests.
SourceE. coli K12 cells
Storage BufferThe enzyme is supplied in 30 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.05% (v/v) Tween 20, and 50% (v/v) glycerol.
Uracil-DNA Glycosylase activity

Uracil-DNA Glycosylase activity

Uracil-DNA Glycosylase activity

Uracil-DNA Glycosylase activity.


References

  1. T. Lindahl et al., DNA N-Glycosidases. J. Biol. Chem. 252(10), 3286-3294 (1977).
  2. M. C. Longo et al., Use of uracil DNA glycosylase to control carry-over contamination in polymerase chain reactions. Gene. 93, 125-128 (1990).
  3. P. Vanghan, T. V. McCarthy, A novel process for mutation detection using uracil DNA glycosylase. Nucleic Acids Res. 26, 810-815 (1998).
  4. T. A. Kunkel, Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc. Natl. Acad. Sci. USA. 82, 488-492 (1985).
  5. P. R. Devchand, et al., Uracil-DNA glycosylase as a probe for protein-DNA interactions. Nucleic Acids Res. 21, 3437-3443 (1993).
  6. P. M. Booth 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).
  7. G. Serrano-Heras et al., Protein p56 from the Bacillus subtilis phage phi29 inhibits DNA-binding ability of uracil-DNA glycosylase. Nucleic Acids Res. 13, 1-9 (2007).

Protocols and Product Manuals

pdf   Uracil-DNA Glycosylase - Product Information