| Recognition site | GAG↑CTC
CTC↓GAG |
| Source | Escherichia coli ICR |
| Assayed on | Lambda DNA (HindIII-digest) |
| Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (HindIII-digest) in 1 hour at 37°C in a total reaction volume of 50 μl. |
| Optimal SE-buffer | G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.) + BSA |
| Enzyme activity (%) | | B | G | O | W | Y | R | | 75 - 100 | 100 | 0 - 10 | 0 - 10 | 75 - 100 | 5 |
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| Optimal temperature | 37oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol; Store at -20°C. |
| Ligations | After 5-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 10 u.a. of enzyme for 16 hours at 37°C.
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| Reagents Supplied with Enzyme |
10 X SE-buffer G, BSA |
| Methylation sensitivity | not tested |
| 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.
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| MSDS: | Download MSDS as PDF |
| References: | Sugisaki H., Maekawa Y., Kanazawa S., Takanami M. Bull. Inst. Chem. Res. Kyoto Univ. 60: 328-335 (1982).
Yankovsky N.K., Fonstein M.Yu., Lashina S.Yu., Bukanov N.O., Yakubovich N.V., Ermakova L.M., Rebentish B.A., Janulaitis A., Debabov V.G. Gene 81: 203-210 (1989).
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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
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