Recognition site | A↑CCWGGT TGGWCC↓A |
Source | Microbacterium arborescens SE |
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 | W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) + BSA |
Enzyme activity (%) | B | G | O | W | Y | Rose | 25 - 50 | 50 - 75 | 75 - 100 | 100 | 50 - 75 | 100 |
|
Optimal temperature | 37oC |
Storage conditions | 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA, 50% glycerol; Store at -20°C. |
Ligation | 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 W, BSA |
Methylation sensitivity | Not blocked by overlapping Dcm methylation (CmCWGG) ACCWGGT. |
Inactivation 20 minutes under | 65oC |
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.
|
Quality control | Restriction Endonucleases: Quality Control |
MSDS: | Download MSDS as PDF |
References: | Nadeev, A.N., Tsaplin, M.M., Abdurashitov, M.A., Dedkov, V.S., Kashirina, J.G., Popichenko, D.V., Degtyarev, S.K. Unpublished observations (2001).
|