| Recognition site | GGCCNNNN↑NGGCC
CCGGN↓NNNNCCGG |
| Source | E.coli strain, that carries the cloned gene Sfi I from Streptomyces fimbriatus |
| Assayed on | T7 DNA |
| Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA in 1 hour at 50°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 | 25 - 50 | 25 - 50 | 25 - 50 | 75 |
|
| Optimal temperature | 50oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C. |
| Ligations | After 10-fold overdigestion with enzyme more than 70% of the DNA fragments can be ligated and recut. In the presence of 10% PEG ligation is better. |
| Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of T7 DNA with 20 u.a. of enzyme for 16 hours at 50°C.
|
| Reagents Supplied with Enzyme |
10 X SE-buffer G, BSA |
| Methylation sensitivity | Blocked by overlapping Dcm methylation (CmCWGG): GGCCWGGNNGGCC.
Not blocked by overlapping Dcm methylation (CmCWGG): GGCCNNNNNGGCCWGG. |
| 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. |
| MSDS: | Download MSDS as PDF |
| References: | Qiang, B.-Q., Schildkraut, I. Nucleic Acids Res. 12: 4507-4515 (1984).
 Dedkov V.S., Degtyarev S. Kh. The resctriction endonucleases detection in colonies of microorganisms Streptomyces and Nocardia // APPLIED BIOCHEMISTRY and MICROBIOLOGY (Russia) Vol 28, 1992, #. 2, 309- 313
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