|Source||Bacillus stearothermophilus 31|
|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 55°C in a total reaction volume of 50 μl.|
|Optimal SE-buffer||O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) + BSA|
|Enzyme activity (%)|
|25 - 50||75 - 100||100||75 - 100||25 - 50||40|
|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 more than 90% of DNA fragments can be ligated. Of these 80% can be recut.|
|Non-specific hydrolisis||No nonspecific activity was detected after incubation of 1 μg of T7 DNA with 10 u.a. of enzyme for 16 hours at 55°C.
|Reagents Supplied with Enzyme||
10 X SE-buffer O, BSA
|Methylation sensitivity||Blocked by overlapping Dcm-methylation (CmCWGG): GAGACCWGG|
Not blocked by methylation GGTCT(5mC)
|Inactivation 20 minutes under||80oC|
|Notes||To obtain 100% activity, BSA should be added to the 1x 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|References:||Shinkarenko, N.M., Lebedeva, N.A., Dedkov, V.S., Abdurashitov, M.A., Degtyarev, S.K. Unpublished observations (1999). |