| Recognition site | RGCGC↑Y
Y↓CGCGR |
| Source | Bacillus stearothermophilus H2 |
| 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 65°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.) + BSA |
| Enzyme activity (%) | | B | G | O | W | Y | Rose | | 50 - 75 | 50 - 75 | 0 - 10 | 10 - 25 | 100 | 100 |
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| Optimal temperature | 65oC |
| Storage conditions | 10 mM KH2PO4(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 10-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 20 u.a. of enzyme for 16 hours at 65°C.
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| Reagents Supplied with Enzyme |
10 X SE-buffer Y, BSA |
| Methylation sensitivity | not tested |
| Inactivation 20 minutes under | 80oC |
| Notes | High enzyme concentration may result in star activity
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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| Quality control | Restriction Endonucleases: Quality Control |
| MSDS: | Download MSDS as PDF |
| References: | Abdurashitov, M.A., Kileva, E.V., Shevchenko, A.V., Degtyarev, S.K.
Unpublished observations (1994).
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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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