Bmu I
| Enzyme name | Bmu I | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Prototype | BfiI | ||||||||||||
| Known analogs | BfiI, BmrI | ||||||||||||
| SKU | SE-E487 | ||||||||||||
| Turbo version | Not available | ||||||||||||
| High-concentration version | Not available | ||||||||||||
| Restriction type | Type IISiType IIS enzymes recognize asymmetric DNA sequences and cleave outside the recognition site at a defined distance. They are widely used in Golden Gate assembly. | ||||||||||||
| Recognition site | 5'… ACTGGG(N)5▼ …3' 3'… TGACCC(N)4▲ …5' | ||||||||||||
| Source | Bacillus megaterium S87 | ||||||||||||
| Optimal buffer | SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM 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 Lambda DNA (HindIII-digest) in 1 hour at 37°C in a total reaction volume of 50 μl. | ||||||||||||
| Assayed on | Lambda DNA (HindIII-digest) | ||||||||||||
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol. Store at -20°C | ||||||||||||
| Ligation | After 2-fold overdigestion with enzyme about 75% of the DNA fragments can be ligated and 95% may be recut. | ||||||||||||
| Nonspecific hydrolysis | Overnight digest with BmuI is not recommended. A 50 μl reaction containing 1 μg of Lambda DNA and 0.5 units of enzyme incubated for 4 hours resulted in the same pattern of DNA bands as a reaction incubated for 1 hour. | ||||||||||||
| Methylation sensitivity | not tested | ||||||||||||
| Reagents supplied with enzyme | 10 X SE-buffer Y | ||||||||||||
| Notes | Enzyme is active in presence of EDTA. | ||||||||||||
| References | No publications found. |