Smi I

Enzyme name Smi I
Prototype SwaI
Known analogs SwaI
SKU SE-E225
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'… ATTT▼AAAT …3'
3'… TAAA▲TTTA …5'
Source Streptococcus milleri S
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 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
2525100752510
Unit definition One unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA (SspI-digest) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on T7 DNA (SspI-digest)
Storage conditions 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligation After 20-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 T7 DNA with 40 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer O, BSA
Notes To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml. Do not use BSA for long incubation.
References
  1. V.S. Dedkov, T.S. Bondar, A.V. Shevchenko, S.Kh. Degtyarev Novel restriction endonuclease SmiI from Streptococcus milleri recognize 5′-ATTT^AAAT-3′. // Molecular Biology and Virusology, No.1, 23-27 (2000). (In Russian)
  2. V.S. Dedkov, S.Kh. Degtyarev Actinobacillus and Streptococcus: producers of isoschizomers of the restriction endonucleases R.Hph I, R.Sau I, R.Nhe I, R.Mbo I and R.Swa I. // Biol. Chem., Vol. 379, 573-574, (1998).
// Код для некролога