Aox I

Recognition Sequence:
↑PuG(5mC)Py

Py(5mC)GPu↓

Source: Arthrobacter oxydans 25K
The enzyme cleaves C5-methylated DNA and does not cut unmodified DNA.

Supplied in: 10 mM KH2PO4 (pH 7.4); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol.

Reaction Conditions:

1X SEBuffer Aox I
Incubate at  60°C

1X SEBuffer Aox I  (pH 8.5 @ 25ºC)

10 mM Tris-HCl;200 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol

Warranty period for the enzyme storage at -20˚C is one year from the date of the last assay indicated on the enzyme vial.

Unit Definition: One unit is defined as the amount of enzyme required to hydrolyze in 1μg of linearized plasmid pMHaeIII/DriI in 1 hour at 60°C in a total reaction volume of 50μl.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`.

Enzyme Properties

Activity in SEBuffers:

SEBuffer B       75-100%

SEBuffer G       50-75%

SEBuffer O       10-25%

SEBuffer W       10-25%

SEBuffer Y            100%

SEBuffer ROSE      50%

When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.

Heat Inactivation: Yes   (65°C for 20 minutes)

Reagents Supplied with Enzyme: 10XSEBuffer Y, BSA (10 mg/ml)