Fig. 4

Digestion properties of recombinant Cac824II as compared to commercial AcuI (New England Biolabs). 250 ng of unmethylated pCONS2.1 plasmid were incubated for 1 h at 37 °C in a reaction volume of 20 µL containing 50 mM potassium acetate, 20 mM Tris–acetate (pH 7.9), 10 mM Magnesium acetate, 100 µg/mL BSA and 0.04 mM S-adenosyl-methionine with (A) No enzyme, (B) purified Cac824II (50 µg), (C) AcuI (5U), and (D) purified Cac824II (50 µg) + AcuI (5U). Lanes M, 1 kb DNA ladder (0.5–10 kbp, NEB). Reactions products were electrophoresed on a 0.8 % agarose gel. An arrow indicates the incomplete digestion product remaining after incubation with Cac824II