Product info: Aox I


Name
Aox I
Cat. #E569E570
Package, u.a.50250
Concentration, u.a./ml500500

Recognition site
↑PuG(5mC)Py
Py(5mC)GPu↓
SourceArthrobacter oxydans 25K
Substrate specificityThe enzyme cleaves C5-methylated DNA and doesn't cut unmodified DNA.
Assayed onDNA 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 definitionOne 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.
AoxI activity assay on DNA pMHaeIII/DriI
Optimal SE-bufferSE-buffer AoxI (10 mM Tris-HCl (pH 7,5 at 25°C); 200 mM KCl; 0,1 mM EDTA, 7 mM 2-mercaptoethanol; 200 μg/ml BSA, 50% glycerol.)
Enzyme activity (%)
BGOWYRose
75 - 10025 - 5010 - 2525 - 5075 - 100100
Reaction bufferSE-buffer AoxI, (10 mM Tris-HCl (pH 7,5 at 25°C); 200 mM KCl; 0,1 mM EDTA, 7 mM 2-mercaptoethanol; 200 μg/ml BSA, 50% glycerol.)
Optimal temperature
60oC
Storage conditions10 mM Tris-HCl (pH 7.4); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethan. Store at -20°C.
Non-specific hydrolisisNo 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 sensitivityThe enzyme cleaves C5-methylated DNA and doesn't cut unmodified DNA.
Inactivation 20 minutes
No
MSDS:Download MSDS as PDF
References:1. Chernukhin V.A., Belichenko O.A., Tarasova M.V., Gonchar D.A., Akishev A.G., Dedkov V.S., Mikhnenkova N.A., Degtyarev S.Kh. Bacterial strain Arthrobacter oxydans - producer of AoxI site specific endonuclease. // Russian Federation patent RU 2399663 C1 (2009).