|Source||An 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 (%)|
|75 - 100||100||25 - 50||25 - 50||25 - 50||75|
|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.|
|Ligation||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||Incubation at 37°C rusults in 75-100% 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.
|Quality control||Restriction Endonucleases: Quality Control|
|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