Thermo Scientific™ Esp3I (BsmBI)
The Esp3I (BsmBI) restriction enzyme recognizes CGTCTC(1/5)^ sites and cuts best at 37°C in Tango (+DTT) buffer (Isoschizomers: BsmBI).
Manufacturer: Thermo Scientific™ ER0451
Lambda DNA digested with Esp3I (BsmBI), 1% agarose, 14 cleavage sites
conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.
5'...C G T C T C (N)1▵...3'
3'...G C A G A G (N)5▵...5' Conditions for 100% Activity:
- [1X Buffer Tango] + DTT: [33mM Tris-acetate (pH 7.9 at 37°C), 10mM Mg-acetate, 66mM K-acetate and 0.1mg/mL BSA] + 1.0mM DTT
- Incubate at 37°C
- Esp3I is supplied in: 10mM potassium phosphate (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.5mg/mL BSA and 50% (v/v) glycerol
Ligation and Recleavage:
- After 50-fold overdigestion with Esp3I, more than 95% of the DNA fragments can be ligated and recut
- Dam: never overlaps-no effect
- Dcm: never overlaps-no effect
- CpG: completely overlaps-blocked
- EcoKI: never overlaps-no effect
- EcoBI: may overlap-effect not determined
Digestion of Agarose-embedded DNA:
- Minimum 5U of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16hours
The enzyme requires DTT. Freshly made DTT should be added to the reaction buffer. Esp3I cleaves downstream of its recognition site and can generate any desired 4 base 5'-overhangs. This feature is useful for PCR product cloning.