|Source||E.coli strain, that carries the cloned gene SspI from Sphaerotilus species|
|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 37°C in a total reaction volume of 50 μl.|
|Optimal SE-buffer||K (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM KCl; 1 mM DTT.) + BSA|
|Enzyme activity (%)|
|75 - 100||50 - 75||25 - 50||50 - 75||75 - 100||100|
|Storage conditions||10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol. Store at -20°C.|
|Ligation||After 10-fold overdigestion with enzyme 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 37°C.
|Reagents Supplied with Enzyme||
10 X SE-buffer K, BSA|
|Methylation sensitivity||Blocked by methylation AmATATT.|
|Inactivation 20 minutes under||65oC|
|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|References:||Schildkraut, I., Grandoni, R. unpublished observations.|
Chernukhin V.A, Abdurashitov M.A., Tomilov V.N., Gonchar D.A., Degtyarev S.Kh. Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 4, pp 19-27, 2007
|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