Btr I, a novel restriction endonuclease, recognizes the non-palindromic sequence 5′-CACGTC-3′ (-3/-3).
// Nucleic Acids Research, Vol. 28, No 11.(2000).
Btr I
| Enzyme name | Btr I | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Prototype | BtrI | ||||||||||||
| Known analogs | BmgBI | ||||||||||||
| SKU | SE-E277 | ||||||||||||
| Turbo version | Not available | ||||||||||||
| High-concentration version | Not available | ||||||||||||
| Recognition site | 5'… CAC▼GTC …3' 3'… GTG▲CAG …5' | ||||||||||||
| Source | Bacillus stearothermophilus SE-U62 | ||||||||||||
| Optimal buffer | SE-buffer O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) | ||||||||||||
| Optimal temperature | 60 °C | ||||||||||||
| Inactivation temperature | 80 °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 in 1 hour at 60°C in a total reaction volume of 50 μl. | ||||||||||||
| Assayed on | Lambda DNA | ||||||||||||
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; and 50% glycerol; Store at -20°C. | ||||||||||||
| Ligation | After 3-fold overdigestion with enzyme 80% of DNA fragments can be ligated. Of these 90% can be recut. | ||||||||||||
| Nonspecific hydrolysis | No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 5 u.a. of enzyme for 16 hours at 60°C. | ||||||||||||
| Methylation sensitivity | not tested | ||||||||||||
| Reagents supplied with enzyme | 10 X SE-buffer O, BSA | ||||||||||||
| Notes | High enzyme concentration may result in star activity. To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml. Do not use BSA for long incubation. | ||||||||||||
| References |