50 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 100 mM NaCl and 0.1 mg/ml BSA.
Incubate at 30°C.
Ligation and Recleavage
After 50-fold overdigestion with SmiI, more than 80% of the DNA fragments can be ligated in a reaction mixture containing 20-40 u of T4 DNA Ligase/1 µg of fragments and 10% PEG. More than 95% of these can be recut.
Storage Buffer
SmiI is supplied in:
10 mM Tris-HCl (pH 7.4 at 25°C), 200 mM NaCl, 1 mM DTT, 1 mM EDTA, 0.2 mg/ml BSA and 50% (v/v) glycerol.
Digestion of Agarose-embedded DNA
Minimum 10 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded Adenovirus-2 DNA in 16 hours.
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