Bst2B I

Enzyme name Bst2B I
Prototype BsiI
Known analogs BauI, BsiI, BssSI
SKU SE-E043
Turbo version Not available
High-concentration version Not available
Recognition site
5'… C▼TCGTG …3'
3'… GAGCA▲C …5'
Source Bacillus stearothermophilus 2B
Optimal buffer SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature 60 °C
Inactivation temperature 80 °C
Buffer activity
BGOWYROSE
75251025100100
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); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol. Store at -20°C.
Ligation After 10-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 Y, BSA
Notes To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml. Do not use BSA for long incubation.
References
  1. Unpublished data Abdurashitov, M.A., Belichenko, O.A., Degtyarev, S.K Unpublished observations (1995).
  2. V.S. Dedkov, N.I. Rechkunova, S.Kh. Degtyarev, P.A. Zhilkin, A.A. Kolykhalov, V.A. Verkhozina Bacillus sphaericus - a strain producer of restriction endonuclease BsiI. // Soviet Union patent SU 1784642 (1992). (In Russian)
  3. S.Kh. Degtyarev, A.A. Kolykhalov, N.I. Rechkunova, V.S. Dedkov, P.A. Zhilkin BsiI - novel unusual restriction endonuclease. // Molecular Biology(Moscow), Vol.24, No. 1, 244-247 (1990). (In Russian)
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