Figure 3.

Characteristics of nuclease activity of KmAgo. (A) DNA-guided DNA cleavage by KmAgo with various divalent cation cofactors. The reaction was performed for 10 min at 37°C. The fraction of cleaved target for each guide-target combination is shown. (B) Temperature dependence of the cleavage reaction with various combinations of guide and target nucleic acids. (C) Dependence of the efficiency of DNA cleavage on the length of guide DNA. The reaction was performed for 4 min at 37°C. (D) The requirement of the 5′-phosphate group in guide DNA for precise target DNA cleavage. The reaction was performed for 3 or 10 min at 37 or 60°C. The marker (M) lane contains DNA fragments corresponding to the canonical cleavage site. For each experiment, representative gels and means and standard deviations from three to four independent measurements are shown.