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

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.