SacI endonuclease recognizes sequence GAGCT^C, and cuts at C^TCGAG. Enzyme storage solution consists of 10mM Tris-HCl (pH7.4 at 25℃), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol. 1X Buffer SacI is composed of 10mM Bis-Tris Propane-HCl (pH6.5 at 37℃), 10mM MgCl2, 0.1mg/ml BSA. 1X Buffer Y is composed of 33mM Tris-acetate (pH7.9 at 37℃), 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. Enzymatic cleavage and connection efficiency: 50-fold excess of this endonuclease digested for 1 hour, >95% of the fragments cleaved by the enzyme can be connected and re-cut (recut). The active unit is defined as 1 unit of enzyme that completely degrades 1 microgram of λ DNA in a reaction system of 37℃ and 50 microliters.
SacI is an imported restriction endonuclease supplied in 500U units. The product sheet lists the packaging, recognized DNA sequence (GAGCT^C / C^TCGAG), and basic buffer compatibility. Storage solution and buffer compositions are given in detail, and the activity unit is defined.
The manual explains how to store the enzyme at -20 °C, safety precautions for handling, and provides step‑by‑step protocols for single‑enzyme digests as well as double or multiple‑enzyme digests, including recommended reaction volumes, incubation temperatures, and buffer usage. This manual is written in Chinese.
| Recognized DNA sequence | GAGCT^C / C^TCGAG |
|---|---|
| Enzyme concentration | 10 U/µl |
| 1X Buffer SacI composition | 10 mM Bis‑Tris Propane‑HCl (pH 6.5 at 37 °C), 10 mM MgCl₂, 0.1 mg/ml BSA |
SacI recognizes the sequence GAGCT^C (or its complement C^TCGAG).
Use 1X Buffer SacI, which contains 10 mM Bis‑Tris Propane‑HCl (pH 6.5 at 37 °C), 10 mM MgCl₂, and 0.1 mg/ml BSA.