| Recognition site | GATNN↑NNATC
CTANN↓NNTAG |
| Source | Bacillus species 8 |
| 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 60°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.) |
| Enzyme activity (%) | | B | G | O | W | Y | Rose | | 25 - 50 | 100 | 75 - 100 | 75 - 100 | 50 - 75 | 100 |
|
| Optimal temperature | 60oC |
| 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. |
| Ligation | After 5-fold overdigestion with enzyme about 80% of the DNA fragments can be ligated and recut. |
| Non-specific hydrolisis | Long incubation is not recommended. Extended incubations and high concentration of the enzyme may result in star activity.
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| Reagents Supplied with Enzyme |
10 X SE-buffer G |
| Methylation sensitivity | Blocked by overlapping Dam-methylation (GmATC): 5’-GmATCN^NNATC-3’/3’-CTmAGN^NNTAG.
Blocked by overlapping (5mC)-methylation: 5’-GATNN^NNAT(5mC)-3’/3’-(5mC)TANN^NNTAG-5’.
Cuts hemimethylated sequence: 5’-GATNN^NNAT(5mC)-3’/3’-CTANN^NNTAG-5’.
|
| Inactivation 20 minutes under | 80oC |
| Notes | High enzyme concentration (more than 5-fold overdigestion on 1 μg substrate) may result in star activity. |
| Quality control | Restriction Endonucleases: Quality Control |
| MSDS: | Download MSDS as PDF |
| References: | Repin, V.E., Serov, G.D., Puchkova, L.I., Tereshenko, T.A., Lebedev, L.R., Chigikov, V.E. Bioorg. Khim. 19: 583-585 (1993).
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