Unpublished data
Unpublished observations (1998).
AspLE I
| Enzyme name | AspLE I | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Prototype | HhaI | ||||||||||||
| Known analogs | BstHH I, CfoI, HhaI, Hin6I, HinP1I, HspA I | ||||||||||||
| SKU | SE-E221 | ||||||||||||
| 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'… GCG▼C …3' 3'… C▲GCG …5' | ||||||||||||
| Source | Arthrobacter species LE3860 | ||||||||||||
| 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 | не инактивируется | ||||||||||||
| 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 37°C in a total reaction volume of 50 μl. | ||||||||||||
| Assayed on | Lambda DNA | ||||||||||||
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol. Store at -20°C. | ||||||||||||
| Ligation | After 10-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut. | ||||||||||||
| Nonspecific hydrolysis | No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 37°C. | ||||||||||||
| Methylation sensitivity | Blocked by 5'-G(5mC)GC-3'/3-CG(5mC)G-5' methylation. Not blocked by 5'-GCG(5mC)-3'/3`-(5mC)GCG-5` or 5'-GCG(5mC)-3'/3'-CGCG-5'methylation. Cut hemimethylated site: 5'-G(5mC)GC-3'/3'-CGCG-5' | ||||||||||||
| Reagents supplied with enzyme | 10 X SE-buffer O | ||||||||||||
| References |