|Source||Bacillus stearothermophilus Fl|
|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||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 (%)|
|25 - 50||25 - 50||10 - 25||25 - 50||100||50|
|Storage conditions||10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol; Store at -20°C.|
|Ligations||After 3-fold overdigestion with enzyme 90% of DNA fragments can be ligated and 95% of these can be recut.|
|Non-specific hydrolisis||No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 2 u.a. of enzyme for 16 hours at 37°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.|
Long incubation with BSA is not recommend.
To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml.
There is DNA-methyltransferase activity in presence of SAM. It is maximum at 48°C. In presence of 10mM MgCl2 enzyme both modifies and hydrolyzes DNA. If MgCl2 is absent enzyme modifies DNA only. And that DNA become proof against BslFI.
BslF I also cleaves the sequence GGGAC(11/15).
|Quality control||Restriction Endonucleases: Quality Control|
|MSDS:||Download MSDS as PDF|
|References:||Nadeev, A.N., Kashirina, J.G., Nayakshina, T.N., Dedkov, V.S., Mezentseva, N.V., Tomilova, J.E., Degtyarev, S.K.;
Unpublished observations. 2004|
|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