Smi I

Description

Recognition site and hydrolysis position:

Source: Streptococcus milleri S
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)
Optimal SE-buffer: SE-buffer O
Enzyme activity (%):

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.
Ligations:
After 20-fold overdigestion with enzyme >95% of the DNA fragments can be ligated and recut. Ligation >95% in presence of 10% PEG.
Non-specific hydrolisis:
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. Reagents Supplied with Enzyme 10 X SE-buffer O, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
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:
Dedkov, V.S., Bondar, T.S., Shevchenco, A.V., Degtyarev, S.Kh. Mol. Gen. Mikrobiol. Virusol. 1: 23-27 (2000).