Analysis of Specificity of DNA-Methyltransferase M.AspS9I in Cell Lysate by means of Restriction Endonucleases Blocking. // Biotekhnologia (Moscow), No 4, p.30-39 (2009). (In Russian)
AspS9 I
| Enzyme name | AspS9 I | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Prototype | Sau96I | ||||||||||||
| Known analogs | AsuI, Cfr13I, PspPI, Sau96I | ||||||||||||
| SKU | SE-E117 | ||||||||||||
| 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'… G▼GNCC …3' 3'… CCNG▲G …5' |
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| Source | Arthrobacter species S9 | ||||||||||||
| Optimal buffer | SE-buffer W (10 mM Tris-HCl (pH 8.5 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 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); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C. | ||||||||||||
| Ligation | After 20-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 30 u.a. of enzyme for 16 hours at 37°C. | ||||||||||||
| Methylation sensitivity | Bloked by overlapping Dcm methylation(C m CWGG) GGN CC WGG | ||||||||||||
| Reagents supplied with enzyme | 10 X SE-buffer W | ||||||||||||
| Notes | |||||||||||||
| References |