Recognition site:
CAYNN↑NNRTG
GTRNN↓NNYAC
Source: Sphingobacterium mizutae M
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 37°C in a total reaction volume of 50 μl.
Optimal SE-buffer: W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Activity in SEBuffers:
SEBuffer B 10 - 25%
SEBuffer G 10 - 25%
SEBuffer O 75 - 100%
SEBuffer W 100%
SEBuffer Y 10 - 25%
SEBuffer ROSE 60%
When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Optimal temperature: 37°C
Storage conditions: 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; and 50% glycerol.
Store at: -20°C.
Ligation: After 5-fold overdigestion with enzyme 90% 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 10 u.a. of enzyme for 16 hours at 37°C.
Reagents supplied with enzyme: 10 X SE-buffer W
Methylation sensitivity: not tested
Inactivation: 20 minutes under 65°C
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