|Source||Pseudomonas stutzeri N2|
|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 30°C in a total reaction volume of 50 μl.|
|Optimal SE-buffer||Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA|
|Enzyme activity (%)|
|10 - 25||10 - 25||0 - 10||0 - 10||100||30|
|Storage conditions||10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA, 50% glycerol. Store at -20°C.|
|Ligations||After 2-fold overdigestion with enzyme more than 70% of DNA fragments can be ligated. Of these 80% can be 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 2 u.a. of enzyme for 16 hours at 30°C.
|Reagents Supplied with Enzyme||
10 X SE-buffer Y, BSA|
|Methylation sensitivity||not tested|
|Inactivation 20 minutes under||65oC|
|Notes||High enzyme concentration may result in star activity. |
Incubation at 37°C results in 20% 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:||Dedkov, V.S., Kileva, E.V., Abdurashitov, M.A., Gonchar, D.A., Popichenko, D.V., Degtyarev, S.K.
Unpublished observations (2001). |