1X Buffer Tango™: 33mM Tris-acetate (pH 7.9 at 37°C), 10mM Mg-acetate, 66mM K-acetate and 0.1mg/mL BSA
Incubate at 37°C
Bpu1102I is supplied in:10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol
Ligation and Recleavage:
After 50-fold overdigestion with Bpu1102I, more than 80% of the DNA fragments can be ligated in a reaction mixture containing 20-40 u of T4 DNA Ligase/1μg of fragments and 10% PEG. More than 95% of these can be recut
Dam: never overlaps — no effect
Dcm: never overlaps — no effect
CpG: may overlap — no effect.
EcoKI: never overlaps — no effect.
EcoBI: may overlap — effect not determined
Digestion of Agarose-embedded DNA:
Minimum 5units of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16hours