|Source||Bacillus stearothermophilus BA|
|Assayed on||Lambda DNA|
|Unit definition||One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 65°C in a total reaction volume of 50 μl.|
|Optimal SE-buffer||W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) + BSA|
|Enzyme activity (%)|
|25 - 50||25 - 50||75 - 100||100||25 - 50||50|
|Storage conditions||10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 10 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.|
|Ligations||After 10-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.|
|Non-specific hydrolisis||No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 65°C.
|Reagents Supplied with Enzyme||
10 X SE-buffer W, BSA|
|Methylation sensitivity||Blocked by CG methylation.|
|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|References:||Abdurashitov, M.A., Shinkarenko, N.M., Dedkov, V.S., Shevchenko, A.V., Degtyarev, S.K. Unpublished observations (1996). |
|Theoretical diagrams of DNA digestion by this enzyme for the most known DNA substrates:|
To view the fragments length values please point mouse cursor over diagram
M - ladder, 1 - Adeno-2 DNA, 2 - Lambda DNA, 3 - T7 DNA, 4 - pUC19, 5 - pBR322