BstEN I

Enzyme name BstEN I
Prototype EcoNI
Known analogs EcoNI, XagI
SKU SE-E103
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'… CCTNN▼NNNAGG …3'
3'… GGANNN▲NNTCC …5'
Source Bacillus stearothermophillus EM
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 65 °C
Inactivation temperature 80 °C
Buffer activity
BGOWYROSE
50502525100100
Unit definition One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 65°C in a total reaction volume of 50 μl.
Assayed on Lambda DNA
Storage conditions 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligation After 5-fold overdigestion with enzyme about 60% of the DNA fragments can be ligated and recut. Of these 90% can be recut
Nonspecific hydrolysis No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 65°C
Methylation sensitivity Not blocked by overlapping Dcm methylation(C m CWGG) CCTGG NNNAGG, CCTNNN CCAGG.
Reagents supplied with enzyme 10 X SE-buffer Y
Notes
References
  1. Unpublished data Abdurashitov, M.A., Dedkov, V.S., Kileva, E.V., Popichenko, D.V., Degtyarev, S.K Unpublished observations (2000).
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