BstDE I

Enzyme name BstDE I
Prototype DdeI
Known analogs DdeI, HpyF3I
SKU SE-E227
Turbo version 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▼TNAG …3'
3'… GANT▲C …5'
Source Bacillus stearothermophilus DE
Optimal buffer SE-buffer G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)
Optimal temperature 60 °C
Inactivation temperature 80 °C
Buffer activity
BGOWYROSE
75100255010100
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 NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol; Store at -20°C.
Ligation After 20-fold overdigestion with enzyme 90% 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 40 u.a. of enzyme for 16 hours at 60°C.
Methylation sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer G
Notes At 37°C activity is 50%-75% from maximum.
References
  4. Unpublished data Shinkarenko, N.M., Shevchenko, A.V., Dedkov, V.S., Abdurashitov, M.A., Degtyarev, S.K Unpublished observations (1995).
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