BstSF I

Enzyme name BstSF I
Prototype SfeI
Known analogs BfmI, BpcI, SfcI, SfeI
SKU SE-E197
Turbo version Not available
High-concentration version Not available
Restriction type Type IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… C▼TRYAG …3'
3'… GAYRT▲C …5'
Source Bacillus stearothermophilus SF
Optimal buffer SE-buffer O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature 60 °C
Inactivation temperature не инактивируется
Buffer activity
BGOWYROSE
75251005050100
Unit definition One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 60°C in a total reaction volume of 50 μl.
Assayed on Lambda DNA
Storage conditions 10 mM Tris-HCl (pH 7.5); 100 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; and 50% glycerol; Store at -20°C.
Ligation After 3-fold overdigestion with enzyme 95% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysis No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 5 u.a. of enzyme for 16 hours at 60°C.
Methylation sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer O, BSA
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.
References
  2. Unpublished data Belichenko, O.A., Shevchenko, A.V., Dedkov, V.S., Abdurashitov, M.A., Degtyarev, S.K Unpublished observations (1995).
  3. S.Kh. Degtyarev, G.G. Prikhodko, N.I. Rechkunova Substrate specificity determination of restriction endonuclease SfeI. // Bioorg. chem (Moscow), Vol.14, No 6, 848-849 (1988). (In Russian)
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