SmaI endonuclease is an imported packaged product, mainly used for DNA enzymatic cleavage reactions. Its recognition sequence is CCC^GGG, compatible buffers include 1X B, 1X G, 1X O, 1X R, 1X Y and 2X Y, the enzyme cutting temperature is 30℃, inactivation conditions are 65℃, and the enzyme cutting time is 20 minutes. The enzyme cutting efficiency is 50% over the amount of this endonuclease to digest 1 hour, >90% of the fragments cut by the enzyme can be connected and reconnected (recut). Its activity unit is defined as in a 37℃, 50 microliter reaction system, the enzyme amount that completely degrades 1 microgram of λ DNA is defined as 1 activity unit, i.e., 1U.
The document describes the SmaI restriction endonuclease, including its recommended reaction conditions, buffer formulations, and handling procedures. It lists the product codes (e.g., D6633) and provides detailed instructions for setting up digestion reactions, enzyme activity units, and incubation times.
Safety and storage recommendations are also covered, advising storage at -20 °C and handling with disposable gloves. The manual includes buffer recipes such as 10 mM Tris‑HCl (pH 7.4), 100 mM KCl, 1 mM DTT, 1 mM EDTA, 0.2 mg/ml BSA, 50 % glycerol, and a 10 X Buffer Y formulation. This manual is written in Japanese.
| Activity unit definition | 1 unit digests 1 µg DNA in 1 hour at 37 °C |
|---|---|
| Optimal temperature | 23 °C (100 % activity) |
| Standard reaction buffer (10 mM Tris‑HCl) | 10 mM Tris‑HCl (pH 7.4 at 25 °C), 100 mM KCl, 1 mM DTT, 1 mM EDTA, 0.2 mg/ml BSA, 50 % glycerol |
| 10 X Buffer Y composition | 33 mM Tris‑acetate (pH 7.9 at 37 °C), 10 mM magnesium acetate, 66 mM potassium acetate, 0.1 mg/ml BSA |
The enzyme shows 100 % activity at 23 °C; activity drops at higher temperatures.
The 10 X Buffer Y contains 33 mM Tris‑acetate (pH 7.9 at 37 °C), 10 mM magnesium acetate, 66 mM potassium acetate, and 0.1 mg/ml BSA.
The D6633 product provides 300 U of SmaI enzyme.