|Source||Bacillus stearothermophilus H2|
|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||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 (%)|
|50 - 75||50 - 75||0 - 10||10 - 25||100||100|
|Storage conditions||10 mM KH2PO4(pH 7.2); 100 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.|
|Ligations||After 20-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 Y, BSA|
|Methylation sensitivity||not tested|
|Inactivation 20 minutes under||80oC|
|Notes||High enzyme concentration may result in star 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:||Abdurashitov, M.A., Kileva, E.V., Shevchenko, A.V., Degtyarev, S.K.
Unpublished observations (1994). |
|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