Activity in SEBuffers:

SEBuffer B        50 - 75%

SEBuffer G             100%

SEBuffer O       50 - 75%

SEBuffer W      75 -100%

SEBuffer Y       75 -100%

SEBuffer ROSE     100%

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); 50 mM KCl; 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 80% of the DNA fragments can be ligated. Of these, 95% can be 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 55°C.

Reagents supplied with Enzyme: 10 X SE-buffer G, BSA

Methylation sensitivity: Not blocked by overlapping dcm-methylation (CmCWGG): GKGCCCWGG

Blocked by GKG(5mC)MC methylation

Inactivation: 20 minutes under 65°C

Notes: To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml. 

Do not use BSA for long incubation.