Source: Acinetobacter johnsonii R2
Assayed on: Lambda DNA
Unit definition: One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 55°C in a total reaction volume of 50 μl.
Optimal SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Activity in SEBuffers:
SEBuffer B 25 -50%
SEBuffer G 10 -25%
SEBuffer O 10 - 25%
SEBuffer W 25 - 50%
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.
Optimal temperature: 55°C
Storage conditions: 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol;
Store at -20°C.
Ligation: After 3-fold overdigestion with enzyme 95% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis: No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 6 u.a. of enzyme for 16 hours at 55°C.
Reagents Supplied with Enzyme: 10 X SE-buffer Y
Methylation sensitivity: Not blocked by dcm methylation: CCWGG
Inactivation: 20 minutes under 65°C
Notes: At 37°C activity is about 10% from maximum.
* availibility of sample size may be limited
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