| Recognition site | ↑(N)8GACNNNNNNTTYG(N)11↑
↓(N)13CTGNNNNNNAARC(N)6↓ |
| Source | Arthrobacter species NTS |
| Assayed on | T7 DNA |
| Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA in 1 hour at 30°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.) + BSA |
| Enzyme activity (%) | | B | G | O | W | Y | Rose | | 0 - 10 | 0 - 10 | 0 - 10 | 0 - 10 | 100 | 20 |
|
| Optimal temperature | 30oC |
| Storage conditions | 10 mM KH2PO4(pH 7.4); 200 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA, 50% glycerol. Store at -20°C. |
| Ligations | After 3-fold overdigestion with enzyme 70% of DNA fragments can be ligated. Of these 80% can be recut. |
| Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of T7 DNA with 1 u.a. of enzyme for 16 hours at 30°C.
|
| Reagents Supplied with Enzyme |
10 X SE-buffer Y, BSA |
| Methylation sensitivity | not tested |
| Inactivation 20 minutes under | 65oC |
| Notes | To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation. |
| Quality control | Restriction Endonucleases: Quality Control |
| MSDS: | Download MSDS as PDF |