| Recognition site | GG↑CGCGCC
CCGCGC↓GG |
| Source | Pseudomanas alcaligenes BS17 |
| 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.) |
| Enzyme activity (%) | | B | G | O | W | Y | R | | 25 - 50 | 10 - 25 | 0 - 10 | 0 - 10 | 100 | 40 |
|
| Optimal temperature | 37oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 100 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C. |
| Ligations | After 2-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 2 u.a. of enzyme for 16 hours at 37°C.
|
| Reagents Supplied with Enzyme |
10 X SE-buffer Y |
| Methylation sensitivity | Blocked by CG methylation. |
| Inactivation 20 minutes under | 65oC |
| MSDS: | Download MSDS as PDF |
| References: | Dedkov, V.S., Belichenko, O.A., Gonchar, D.A., Akishev, A.G., Yamkovaya, T.V., Mezentseva, N.V., Tomilova, J.E., Abdurashitov, M.A., Degtyarev, S.K.
Unpublished observations(2005)
|
Theoretical diagrams of DNA digestion by this enzyme for the most known DNA substrates:
To view the fragments length values please point mouse cursor over diagram |
M - ladder, 1 - Adeno-2 DNA, 2 - Lambda DNA, 3 - T7 DNA, 4 - pUC19, 5 - pBR322
|
