| Recognition site | TCG↑CGA
AGC↓GCT |
| Source | Nocardia rubra |
| Assayed on | Lambda DNA (dam-) |
| Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dam-) in 1 hour at 37°C in a total reaction volume of 50 μl. |
| Optimal SE-buffer | W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) |
| Enzyme activity (%) | | B | G | O | W | Y | R | | 0 - 10 | 10 - 25 | 75 - 100 | 100 | 10 - 25 | 100 |
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| Optimal temperature | 37oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol; Store at -20°C. |
| Ligations | After 5-fold overdigestion with enzyme about 20% of the DNA fragments can be ligated. In the presence of 10% PEG ligation is better. |
| 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 W |
| Methylation sensitivity | Blocked by overlapping Dam methylation (GmATC): TCGCGATC. |
| Inactivation 20 minutes under | 80oC |
| MSDS: | Download MSDS as PDF |
| References: | Comb, D.G., Schildkraut, I., Greenough, L. Unpublished observations.
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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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