Uracil-DNA Glycosylase
ROCHE/11444646001 - recombinant from E. coli K 12
Synonym: PCR; UDG
Product Type: Chemical
| application(s) | life science and biopharma |
| color | colorless |
| concentration | 1000 U/mL |
| foreign activity | endonuclease 20 units, none detected |
| RNases 10 units, none detected | |
| form | solution |
| manufacturer/tradename | Roche |
| packaging | pkg of 100 U |
| pH | 7.9-8.1 (68 °F) |
| Quality Level | 100  |
| recombinant | expressed in E. coli |
| solubility | water: miscible |
| storage temp. | −20°C |
| suitability | suitable for molecular biology |
| technique(s) | mutagenesis: suitable |
| PCR: suitable |
| 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 |
| 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. |
| Other Notes: | For life science research only. Not for use in diagnostic procedures. |
| Physical form: | The enzyme is supplied as 1U/μl solution in storage buffer. |
| 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. |
| 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. |
| Storage and Stability: | OK to freeze (after DNA synthesis) |
| 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 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | No data available |
| Flash Point(C) | No data available |
| Storage Temp. | −20°C |
| UNSPSC | 41106300 |