Activity in SEBuffers:

SEBuffer B       75 -100%

SEBuffer G       50 -  75%

SEBuffer O         0 -  1 0%

SEBuffer W       10 - 25 %

SEBuffer Y        50 -  75%

SEBuffer ROSE        10%

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); 100 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.

Store at: -20°C. 

Store at: -70°C is recommended for periods longer than 7 days.

Ligation: After 3-fold overdigestion with enzyme > 95% of Ad2 DNA fragments can be ligated with T4 DNA Ligase and recut.

Non-specific hydrolisis: No nonspecific activity was detected after incubation of 1 μg of Ad2 DNA with 4 units of enzyme for 16 hours at 37°C.

Reagents supplied with enzyme: 10 X SE-buffer Rig I, BSA

Methylation sensitivity: Blocked by mCG or GmC methylation:

5`-GGC(m5C)GGCC-3`/3`-CCGG(m5C)CGG-5` or 

5`-GG(m5C)CGG(m5C)C-3`/3`-C(m5C)GGC(m5C)GG-5`

Inactivation: 20 minutes under 65°C

Notes: Do not use BSA for long incubation. To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.