|Catalog number||E479||E480 |
|Package, u.a.||100||500 |
|Concentration, u.a./ml||1000||1000 |
|Methylation sensitivity||not tested|
|Origin||Bacillus stearothermophilus Fl|
|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.|
|Assayed on||Lambda DNA|
|Ligation||After 3-fold overdigestion with enzyme 90% of DNA fragments can be ligated and 95% can be recut.|
|Non-specific activity||No nonspecific activity was detected after incubation of 1 ug of Lambda DNA with 2 u.a. of enzyme for 16 hours at 37°C.|
|Optimum temperature||37 oC|
|Inactivation 20 minutes||80oC|
|Optimum 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|
Note: 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 ug/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).
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Unpublished observations. 2004