| Recognition site | CCTNN↑NNNAGG
GGANNN↓NNTCC |
| Source | Bacillus stearothermophillus EM |
| Assayed on | Lambda DNA |
| Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 65°C in a total reaction volume of 50 μl. |
| Optimal SE-buffer | Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) |
| Enzyme activity (%) | | B | G | O | W | Y | R | | 50 - 75 | 50 - 75 | 25 - 50 | 25 - 50 | 100 | 100 |
|
| Optimal temperature | 65oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 200 μg/ml BSA; 7 mM 2-mercaptoethanol; 50% glycerol. Store at -20°C. |
| Ligations | After 3-fold overdigestion with enzyme about 60% of the DNA fragments can be ligated. Of these 90% can be recut |
| Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 65°C
|
| Reagents Supplied with Enzyme |
10 X SE-buffer Y |
| Methylation sensitivity | Not blocked by overlapping Dcm methylation(CmCWGG): CCTGGNNNAGG,
CCTNNNCCAGG. |
| Inactivation 20 minutes under | 80oC |
| MSDS: | Download MSDS as PDF |
| References: | Abdurashitov, M.A., Dedkov, V.S., Kileva, E.V., Popichenko, D.V., Degtyarev, S.K. Unpublished observations (2000).
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