In the present study, we evaluated the antimicrobial activity of 16 Jordanian medicinal plant extracts against four reference bacteria; Staphylococcus aureus, Enterobacter faecalis, Escherichia coli, and Salmonella typhi. For that purpose, whole plants were extracted and antimicrobial susceptibility testing and minimum inhibitory concentration (MIC) were determined. Ethanolic extracts of most medicinal plants exerted a dose-dependent cytotoxiciy against different reference bacteria. Origanum syriaca, Varthemia iphionoides, Psidium guajava, Sarcopoterium spinosa plant extracts were most active against S. aureus (MIC; 70 μg/mL), E. faecalis (MIC; 130 μg/mL), E. coli (MIC; 153 μg/mL), and S. typhi (MIC; 110 μg/mL), respectively. Results indicate that medicinal plants grown in Jordan might be a valuable source of starting materials for the extraction and/or isolation of new antibacterial agents.