|Source||Streptococcus milleri S|
|Assayed on||T7 DNA (SspI-digest)|
|Unit definition||One unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA (SspI-digest) in 1 hour at 37°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||25 - 50||100||75 - 100||25 - 50||10|
|Storage conditions||10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol. Store at -20°C.|
|Ligation||After 20-fold overdigestion with enzyme >95% of the DNA fragments can be ligated and recut. Ligation >95% in presence of 10% PEG.|
|Non-specific hydrolisis||No nonspecific activity was detected after incubation of 1 μg of T7 DNA with 40 u.a. of enzyme for 16 hours at 37°C.
|Reagents Supplied with Enzyme||
10 X SE-buffer O, BSA
For E225T and E226T an additional 10X SE-buffer ROSE is supplied.
|Methylation sensitivity||not tested|
|Inactivation 20 minutes under||65oC|
|Notes||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.
Turbo Smi I can be used for short time (10 min) DNA digestion as well as for standard reaction. The reaction can be performed using optimal or universal (ROSE) Buffer
Turbo DNA Digestion:
-Fast DNA analysis
-Fast preparation of vectors for cloning
1 μl of Turbo Smi I cuts 1 μg of DNA in 1 x SE-Buffer O or universal 1 x SE-Buffer ROSE in 10 min (see the protocol below). A short time of DNA digestion requires high quality purification of DNA sample. The enzyme may be used in standard conditions of DNA cleavage (E225).
Turbo reaction protocol:
20 μl of the reaction volume:
10 x SE Buffer ROSE - 2 μl
DNA - 0,2-1 μg
Nuclease-free water - to 20 μl
+ 1μl of Turbo Smi I
Incubate at 37°C for 10 min.
|Quality control||Restriction Endonucleases: Quality Control|
|MSDS:||Download MSDS as PDF|
|References:||Dedkov, V.S., Bondar, T.S., Shevchenco, A.V., Degtyarev, S.Kh. Mol. Gen. Mikrobiol. Virusol. 1: 23-27 (2000).|
|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