| Recognition site | ↑PuG(5mC)Py
Py(5mC)GPu↓
|
| Source | Arthrobacter oxydans 25K |
| Substrate specificity | The enzyme cleaves C5-methylated DNA and doesn't cut unmodified DNA.
|
| Assayed on | DNA pMHaeIII/DriI is a linearized plasmid pMHaeIII. pMHaeIII carries a gene of DNA-methyltransferase M.HaeIII, which methylates sites 5`-GGCC-3` producing 5`-GG(5mC)C-3`/3`-C(5mC)GG-5`. |
| Unit definition | One unit is defined as the amount of enzyme
required to hydrolyze
in 1 μg of linearized plasmid pMHaeIII/DriI in 1 hour at 60°C in a total reaction volume of 50 μl. As a result a linearized plasmid pMHaeIII/DriI disappears (see run 4 in the figure). |
AoxI activity assay on DNA pMHaeIII/DriI
Lanes:
1 – Control DNA pMHaeIII/DriI,
2 – 0.5 μl Aox I
3 – 1 μl Aox I
4 – 2 μl Aox I
5 – 1 Kb SE DNA-markers
Products were separated in 1% agarose gel in Buffer TAE. |
 |
| Reaction buffer | SE-buffer AoxI, (10 mM Tris-HCl (pH 8,5 at 25°C); 3 mM MgCl2; 1 mM DTT.) |
| Optimal temperature | 60oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.4); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethan. Store at -20°C. |
| Non-specific hydrolisis | No detectable degradation of 1μg of Lambda DNA was observed after incubation with 1 units of enzyme for 16 hours at 60°C in a total reaction volume of 50 μl.
|
| Reagents Supplied with Enzyme |
10 X SE-buffer AoxI |
| Methylation sensitivity | The enzyme cleaves C5-methylated DNA and doesn't cut unmodified DNA. |
| Inactivation 20 minutes | No |
| MSDS: | Download MSDS as PDF |
