| Recognition site | GGCCGG↑CC
CC↓GGCCGG |
| Source | Rhizobium yangligense |
| Assayed on | Adenovirus -2 DNA |
| Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of Ad2 DNA in 1 hour at 37°C in a total reaction volume of 50 μl. |
| Optimal SE-buffer | SE-buffer RigI (10 mM Tris-HCl (pH 8.5 at 25°C); 5 mM MgCl2; 1mM DTT.) + BSA |
| Enzyme activity (%) | | B | G | O | W | Y | Rose | | 75 - 100 | 50 - 75 | 0 - 10 | 10 - 25 | 50 - 75 | 10 |
|
| Optimal temperature | 37oC |
| 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.
Storage at -20°C.
Storage 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 RigI, 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 | 65oC |
| 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. |
| Quality control | Restriction Endonucleases: Quality Control |
| MSDS: | Download MSDS as PDF |