BstDE I

Enzyme nameBstDE I
PrototypeDdeI
Known analogsDdeI, HpyF3I
SKUSE-E227
Turbo versionAvailable
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▼TNAG …3'
3'… GANT▲C …5'
SourceBacillus stearothermophilus DE
Optimal bufferSE-buffer G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)
Optimal temperature60 °C
Inactivation temperature80 °C
Buffer activity
BGOWYROSE
75100255010100
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 NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol; Store at -20°C.
LigationAfter 20-fold overdigestion with enzyme 90% of the DNA fragments can be ligated and recut
Nonspecific hydrolysisNo 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 sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer G
NotesAt 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 Unpublished data Shinkarenko, N.M., Shevchenko, A.V., Dedkov, V.S., Abdurashitov, M.A., Degtyarev, S.K Unpublished observations (1995).
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