11444646001

Uracil-DNA Glycosylase

recombinant from E. coli K 12

• Synonym(s): PCR, UDG
• UNSPSC Code: 41106300
• NACRES: NA.54
• Recombinant: expressed in E. coli
• Concentration: 1000 U/mL

Properties

• recombinant: expressed in E. coli
• form: solution
• packaging: pkg of 100 U
• manufacturer/tradename: Roche
• concentration: 1000 U/mL
• technique(s): PCR: suitable, mutagenesis: suitable
• color: colorless
• pH: 7.9-8.1 (68 °F)
• solubility: water: miscible
• suitability: suitable for molecular biology
• application(s): life science and biopharma
• foreign activity: RNases 10 units, none detected, endonuclease 20 units, none detected
• storage temp.: −20°C
• Quality Level: 100

Description

General description

Uracil-DNA Glycosylase (UNG) contains the highly active recombinant form of the equally named enzyme found in prokaryotes and eukaryotes. It hydrolyzes uracil-glycosidic bonds in single- or double-stranded DNA, excising uracil and creating alkali-sensitive abasic sites in the DNA. These abasic sites can be hydrolyzed by endonuclease, heat, or alkali treatment. Depending on how the DNA is prepared, Uracil-DNA Glycosylase can be used to achieve general, site-specific, or strand-specific U-DNA cleavage.

Application

Uracil-DNA Glycosylase can be used to cleave DNA at any site where a deoxyuridylate residue has been incorporated. U-DNA can be prepared by in vitro methods like PCR. General, site-specific, or strand-specific cleavage can be achieved with uracil-DNA glycosylase, depending on how the U-DNA is prepared. Uracil-DNA Glycosylase can therefore help you to:

• Prevent carryover contamination in PCR
• Increase the efficiency of site-directed mutagenesis procedures
• Label oligonucleotide probes

Biochem/physiol Actions

• Uracil-DNA glycosylase hydrolyzes uracil-glycosidic bonds at U-DNA sites in single- and doublestranded DNA, excising uracil and creating alkali sensitive abasic sites in the DNA.
• The enzyme is more active on single-stranded DNA than on double-standed DNA.
• Activity was also observed on small U-DNA oligonucleotides and on dUMP (Duncan, unpublished observations).
• Uracil-DNA glycosylase is inactive on RNA and native, uracil-free DNA.

Heat inactivation: 95 °C for 10 min
Uracil-DNA glycosylase remains partially active (<10%) after an incubation period of 30 minutes at 95 °C.

Physical form

The enzyme is supplied as 1U/μl solution in storage buffer.

Preparation Note

OK to freeze (after DNA synthesis)

Analysis Note

Carryover prevention activity is assayed by adding approximately 10⁵dU that contains templates prior to the amplification reaction. After UNG treatment, no amplification products could be detected. The enzyme does not contain any contaminating exo- or endonucleases and is tested for the absence of RNases.

Other Notes

For life science research only. Not for use in diagnostic procedures.

One unit is defined as the amount of Uracil-DNA Glycosylase necessary to completely degrade 1 μg purified single-stranded uracil containing DNA (bacteriophage M13, grown in E.coli CJ 236 dut-ung-) at +37 °C in 60 minutes.
One Lindahl unit is defined as the amount of enzyme necessary to release of 1 μmol uracil at +37 °C in 1 minute. One Lindahl unit is comparable to 520,000 U based on our unit definition.

Volume Activity: 1 U/μl

Safety Information

• Storage Class: 12 - Non Combustible Liquids
• wgk: WGK 1
• flash_point_f: No data available
• flash_point_c: No data available