The function of DNA ligases is to repair nicked DNA strands. One such enzyme, ChV ligase, requires ATP to complete the formation of the phosphodiester bond between the ends of a single-strand break. ChV ligase is the smallest known ATP-dependent ligase whose three-dimensional structure has been solved. By studying its interaction with nonspecific DNA, it is possible to determine the feasibility of the use of ChV ligase for future research. The protein, in its purified form, was combined with DNA, and the fluorescence anisotropies of different samples were measured. The anisotropy data points were fitted to standard equations to determine the dissociation constants of the DNA complexes with ChV ligase in its adenylated and apo states. The resulting dissociation constants, which show that the protein has a high affinity for nonspecific DNA, suggest that ChV ligase is an appropriate enzyme to test in future studies concerning protein translocation on nonspecific DNA.