| Recognition site | GGTCTC(N)1↑
CCAGAG(N)5↓ |
| Source | Bacillus stearothermophilus 31 |
| Assayed on | T7 DNA |
| Unit | One unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA in 1 hour at 55°C in a total reaction volume of 50 μl. |
| Optimal SE-buffer | O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) + BSA |
| Enzyme activity (%) | | B | G | O | W | Y | R | | 25 - 50 | 75 - 100 | 100 | 75 - 100 | 25 - 50 | 40 |
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| Optimal temperature | 55oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol; Store at -20°C. |
| Ligations | After 5-fold overdigestion with enzyme more than 90% of DNA fragments can be ligated. Of these 80% can be recut. |
| Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of T7 DNA with 5 u.a. of enzyme for 16 hours at 55°C.
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| Reagents Supplied with Enzyme |
10 X SE-buffer O, BSA |
| Methylation sensitivity | Not blocked by methylation GGTCTmC |
| Inactivation 20 minutes under | 80oC |
| Notes | To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation. |
| MSDS: | Download MSDS as PDF |
| References: | Shinkarenko, N.M., Lebedeva, N.A., Dedkov, V.S., Abdurashitov, M.A., Degtyarev, S.K. Unpublished observations (1999).
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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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