| Recognition site | GACNNNN↑NNGTC
CTGNN↓NNNNCAG |
| Source | Deinococcus species D2 |
| 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 37°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.) + BSA |
| Enzyme activity (%) | | B | G | O | W | Y | R | | 75 - 100 | 75 - 100 | 25 - 50 | 50 - 75 | 100 | 30 |
|
| Optimal temperature | 37oC |
| 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 5-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and 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 37°C
|
| Reagents Supplied with Enzyme |
10 X SE-buffer Y, BSA |
| Methylation sensitivity | not tested |
| Inactivation 20 minutes under | 80oC |
| 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.
|
| MSDS: | Download MSDS as PDF |
| References: | Abdurashitov, M.A., Kileva, E.V., Dedkov, V.S., Degtyarev, S.K.
Unpublished observations (1996).
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