Recognition site | GGN↑NCC CCN↓NGG |
Source | Pseudomonas species N4 |
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 | Rose | 10 - 25 | 10 - 25 | 10 - 25 | 25 - 50 | 100 | 50 |
|
Optimal temperature | 37oC |
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 10-fold overdigestion with enzyme more than 95% 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 20 u.a. of enzyme for 16 hours at 37°C.
|
Reagents Supplied with Enzyme |
10 X SE-buffer Y |
Methylation sensitivity | Blocked by
5`-GGNN(5mC)C-3`/3-C(5mC)NNGG-5` or
5`-GGNN(5mC)C-3`/3-CCNNGG-5`
methylation |
Inactivation 20 minutes under | 65oC |
Quality control | Restriction Endonucleases: Quality Control |
MSDS: | Download MSDS as PDF |
References: | Abdurashitov, M.A., Kileva, E.V., Shevchenko, A.V., Dedkov, V.S., Degtyarev, S.Kh. Unpublished observations (1995).
Chernukhin V.A, Abdurashitov M.A., Tomilov V.N., Gonchar D.A., Degtyarev S.Kh. Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 4, pp 19-27, 2007
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