Thermo Scientific™ Eco31I (BsaI) (10 U/μL)

Description

Lambda DNA (dcm-) digested with Eco31I (BsaI), 0.7% agarose, 2 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'...G G T C T C (N)₁▵...3'

3'...C C A G A G (N)₅▵...5'

Conditions for 100% Activity:

• 1X Buffer G:10mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl₂, 50mM NaCl and 0.1mg/mL BSA
• Incubate at 37°C

Storage Buffer:

• Eco31I is supplied in:10mM Tris-HCl (pH 7.5 at 25°C), 200mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol

Ligation and Recleavage:

• After 50-fold overdigestion with Eco31I, more than 95% of DNA fragments can be ligated and recut

Methylation Effects:

• Dam: never overlaps-no effect
• Dcm: may overlap-cleavage impaired
• CpG: may overlap-cleavage impaired
• 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 16 hours

Note:

Assayed using lambda DNA (dcm-) (#SD0021), as one of the two Eco31I recognition sites in lambda DNA is difficult to cleave. Eco31I cleavage is impaired by overlappingdcm methylation. To avoiddcm methylation, use adam,dcm- strain such as GM2163 (#M0099). Low salt, high glycerol (>5%) concentrations, pH >8.0 or a large excess of enzyme may result in star activity. Eco31I cleaves downstream of its recognition site and can generate any desired 4 base 5'-overhangs. This feature is useful for direct PCR product cloning.