BstSF I

Enzyme nameBstSF I
PrototypeSfeI
Known analogsBfmI, BpcI, SfcI, SfeI
SKUSE-E197
Turbo versionNot available
High-concentration versionNot available
Restriction typeType 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'
SourceBacillus stearothermophilus SF
Optimal bufferSE-buffer O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature60 °C
Inactivation temperatureне инактивируется
Buffer activity
BGOWYROSE
75251005050100
Unit definitionOne 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 onLambda DNA
Storage conditions10 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.
LigationAfter 3-fold overdigestion with enzyme 95% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisNo 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 sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer O, BSA
NotesTo 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 Unpublished data Belichenko, O.A., Shevchenko, A.V., Dedkov, V.S., Abdurashitov, M.A., Degtyarev, S.K Unpublished observations (1995).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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