Small molecules that interact with DNA are known to be effective as anticancer and antimicrobial agents. Therefore it is significant to search for new molecules interacting with DNA as potential new therapeutic agents. In this study, we aimed to investigate interactions of novel fluorine substituted imine compound with DNA, (E)-2-((3,5-bis(trifluoromethyl) phenylimino)methyl)-4,6-dimethoxyphenol, and investigate its biological activities. DNA interactions of the compound were investigated by UV-Vis absorption spectroscopy and gel electrophoresis. The results demonstrated that the compound binds to DNA via intercalation. Agarose gel electrophoresis experiments showed that the compound does not cleave pBR322 plasmid DNA hydrolytically or oxidatively. Furthermore, mutagenic, anti-mutagenic, and antimicrobial activities of the compound were studied by Ames and broth microdilution test, respectively. The compound showed mutagenic activity on both TA98 and TA100 strains. Also, the antimutagenic activity was observed in TA100 strain of S. typhimurium. It demonstrated antimicrobial activity against the microorganisms tested in the concentration range of 16-64μg/μL. The results show that the compound intercalates with DNA and has promising biological activities.