Thermo Scientific™ Esp3I (BsmBI)

Descrizione

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

Storage Buffer:

• 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

Methylation Effects:

• 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

Note:

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.