Rig I
| Enzyme name | Rig I | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Prototype | FseI | ||||||||||||
| Known analogs | FseI | ||||||||||||
| SKU | SE-E529 | ||||||||||||
| Turbo version | Not available | ||||||||||||
| High-concentration version | Not available | ||||||||||||
| Restriction type | Type IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites. | ||||||||||||
| Recognition site | 5'… GGCCGG▼CC …3' 3'… CC▲GGCCGG …5' | ||||||||||||
| Source | Rhizobium yangligense | ||||||||||||
| Optimal buffer | SE-buffer RigI (10 mM Tris-HCl (pH 8.5 at 25°C); 5 mM MgCl2; 1mM DTT.) | ||||||||||||
| Optimal temperature | 37 °C | ||||||||||||
| Inactivation temperature | 65 °C | ||||||||||||
| Buffer activity |
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| 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. | ||||||||||||
| Assayed on | Adenovirus -2 DNA | ||||||||||||
| 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. | ||||||||||||
| Nonspecific hydrolysis | No nonspecific activity was detected after incubation of 1 μg of Ad2 DNA with 4 units of enzyme for 16 hours at 37°C. | ||||||||||||
| 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` | ||||||||||||
| Reagents supplied with enzyme | 10 X SE-buffer RigI, BSA | ||||||||||||
| 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. | ||||||||||||
| References | No publications found. |