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11444646001 Roche

Uracil-DNA Glycosylase

recombinant from E. coli K 12

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Properties

Related Categories Molecular Biology, Molecular Biology Enzymes, Ultra Pure Modifying Enzymes More...
Quality Level   100
form   solution
packaging   pkg of 100 U
mfr. no.   Roche
shipped in   dry ice
storage temp.   −20°C

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.

Specificity

• 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• Activity was also observed on small U-DNA oligonucleotides and on dUMP (Duncan, unpublished observations).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.

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

Quality

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.

Unit Definition

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

Physical form

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

Other Notes

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

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis (COA)

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Protocols & Articles

Articles

DNA Damage and Repair

Damage to cellular DNA is involved in mutagenesis and the development of cancer. The DNA in a human cell undergoes several thousand to a million damaging events per day, generated by both external (e...
Keywords: AGE, Alkylations, Apoptosis, Biochemistry, Cancer, Carcinogens, Catalysis, Clinical, DNA replication, Deaminations, Degradations, Eliminations, Environmental, Enzymology, Gene expression, Genetic, Infrared spectroscopy, Metabolites, Methylations, Mutagens, Oncology, Oxidations, Polymorphisms, Rearrangements, Recombination, Substitutions, Transcription, transformation

Roche PCR Reagents and PCR Protocols

For additional information on Hot Start PCR technology, protocols, and to download our free Hot Start PCR eBook, please visit our Hot Start PCR resource guide.
Keywords: Amplification, Cloning, Gas chromatography, Gene expression, Polymerase chain reaction, Polymerase chain reaction - quantitative, Purification, Sequencing, Transcription

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