Polymicrobial biofilms pose a significant challenge in clinical settings due to their complex composition and antibiotic resistance. Recent advances in omics technologies have revolutionized the diagnosis and treatment of biofilm-related infections. This chapter discusses the application of omics approaches, including genomics, transcriptomics, proteomics, and metabolomics, in the diagnosis of polymicrobial biofilms and the identification of novel therapeutic targets. We highlight the potential of omics-driven strategies in the development of personalized medicine and the discovery of new antimicrobial agents.