Bso31 I

Enzyme name Bso31 I
Prototype Eco31I
Known analogs BsaI, BspTNI, Eco31I
SKU SE-E285
Turbo version Available
High-concentration version Not available
Restriction type Type IISiType IIS enzymes recognize asymmetric DNA sequences and cleave outside the recognition site at a defined distance. They are widely used in Golden Gate assembly.
Recognition site
5'… GGTCTCN▼ …3'
3'… CCAGAG(N)5▲ …5'
Source Bacillus stearothermophilus 31
Optimal buffer SE-buffer O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature 55 °C
Inactivation temperature 80 °C
Buffer activity
BGOWYROSE
2575100752540
Unit definition One unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA in 1 hour at 55°C in a total reaction volume of 50 μl.
Assayed on T7 DNA
Storage conditions 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligation After 5-fold overdigestion with enzyme more than 90% of DNA fragments can be ligated. Of these 80% can be recut.
Nonspecific hydrolysis No nonspecific activity was detected after incubation of 1 μg of T7 DNA with 10 u.a. of enzyme for 16 hours at 55°C.
Methylation sensitivity Blocked by overlapping Dcm-methylation (CmCWGG) GAGA CC WGG Not blocked by methylation GGTCT(5mC)
Reagents supplied with enzyme 10 X SE-buffer O, 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
  2. Unpublished data Shinkarenko, N.M., Lebedeva, N.A., Dedkov, V.S., Abdurashitov, M.A., Degtyarev, S.K Unpublished observations (1999).
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