Uracil Cleavage System, including Endonuclease VIII (0.375 mL) and Uracil-DNA Glycosylase (0.375 mL).
The Uracil Cleavage System is intended for molecular biology applications. This product is neither intended for the diagnosis, prevention or treatment of a disease, nor has it been validated for such use either alone or in combination with other products.
Features
- Two-enzyme system, Uracil DNA Glycosylase (UDG) and Endonuclease VIII
- UDG catalyzes the excision of the uracil base, creating an abasic site with an intact phosphodiester backbone
- Endonuclease VIII breaks the phosphodiester backbone both 3’ and 5’ to the abasic site, liberating the deoxyribose sugar
Product Details
The Uracil Cleavage System provides two enzymes, which, when added sequentially to a reaction containing a synthetic DNA fragment containing a deoxy-uracil, generate a single nucleotide gap at the location of the uracil residue. The two individual enzyme components, Uracil DNA Glycosylase and Endonuclease VIII, provided at a 10x concentration, are to be added to a reaction containing a uracil-containing polynucleotide sequence. UDG catalyzes the excision of the uracil base, creating an abasic site with an intact phosphodiester backbone (1–2). The lyase activity of Endonuclease VIII breaks the phosphodiester backbone, both 3ʹ and 5ʹ, to the abasic site, liberating the deoxyribose sugar (3–4).
UDG is supplied in 10 mM Tris-HCl, 50 mM NaCl, 1 mM DTT, 0.1 mM EDTA and 50% glycerol; pH 7.5 at 25°C.
Endonuclease VIII is supplied in 10 mM Tris-HCl, 250 mM NaCl, 0.1 mM EDTA and 50% glycerol; pH 8.0 at 25°C.
Performance
| Test | Amount tested | Specification |
| Purity | n/a | >99% |
| Single-stranded exonuclease | 10 μL | <5% released |
| Double-stranded exonuclease | 10 μL | <1% released |
| Double-stranded endonuclease | 10 μL | No conversion |
| E. coli DNA contamination | 5 μL | <10% copies |