Novel restriction endonuclease SmiI from Streptococcus milleri recognize 5′-ATTT^AAAT-3′. // Molecular Biology and Virusology, No.1, 23-27 (2000). (In Russian)
Smi I
| Enzyme name | Smi I | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Prototype | SwaI | ||||||||||||
| Known analogs | SwaI | ||||||||||||
| SKU | SE-E225 | ||||||||||||
| Turbo version | 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'… ATTT▼AAAT …3' 3'… TAAA▲TTTA …5' | ||||||||||||
| Source | Streptococcus milleri S | ||||||||||||
| 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 | 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 T7 DNA (SspI-digest) in 1 hour at 37°C in a total reaction volume of 50 μl. | ||||||||||||
| Assayed on | T7 DNA (SspI-digest) | ||||||||||||
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol. Store at -20°C. | ||||||||||||
| Ligation | After 20-fold overdigestion with enzyme >95% of the DNA fragments can be ligated and recut. | ||||||||||||
| Nonspecific hydrolysis | No nonspecific activity was detected after incubation of 1 μg of T7 DNA with 40 u.a. of enzyme for 16 hours at 37°C. | ||||||||||||
| Methylation sensitivity | not tested | ||||||||||||
| Reagents supplied with enzyme | 10 X SE-buffer O, BSA | ||||||||||||
| Notes | 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 |