|Catalog number||E123||E124||E123X||E124X |
|Package, u.a.||1000||5000||1000||5000 |
|Concentration, u.a./ml||10000||10000||40000||40000 |
|Methylation sensitivity||Blocked by overlapping Dcm methylation (CmCWGG): GGCCWGGNNGGCC.|
Not blocked by overlapping Dcm methylation (CmCWGG): GGCCNNNNNGGCCWGG.
|Origin||E.coli strain, that carries the cloned gene Sfi I from Streptomyces fimbriatus|
|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.|
|Assayed on||T7 DNA|
|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 activity||No nonspecific activity was detected after incubation of 1 ug of T7 DNA with 20 u.a. of enzyme for 16 hours at 50°C.|
|Optimum temperature||50 oC|
|Inactivation 20 minutes||65oC|
|Optimum 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|
Note: To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 ug/ml.
Do not use BSA for long incubation.
References: Qiang, B.-Q., Schildkraut, I. Nucleic Acids Res. 12: 4507-4515 (1984).