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11444646001

Roche

Uracil-DNA Glycosylase

recombinant from E. coli K 12

Synonym(s):

PCR, UDG

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About This Item

UNSPSC Code:
41106300

recombinant

expressed in E. coli

Quality Level

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

Related Categories

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  • Activity was also observed on small U-DNA oligonucleotides and on dUMP (Duncan, unpublished observations).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

Storage and Stability

OK to freeze (after DNA synthesis)

Other Notes

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

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

No data available

Flash Point(C)

No data available


Certificates of Analysis (COA)

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Articles

DNA damage and repair mechanism is vital for maintaining DNA integrity. Damage to cellular DNA is involved in mutagenesis, the development of cancer among others.

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