Ethanolic and aqueous extracts of Lactuca tatarica leaves, roots, and acetonic extract of its leaves were studied for their antimicrobial activity against clinically important standard microbial strains (Candida albicans, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Bacillus anthracoides). For the preparation of the stock solutions, the crude ethanolic and acetonic extracts were dissolved in 100% DMSO, the aqueous extracts were dissolved in distilled water. The concentration of crude extracts was prepared at 150 mg/mL and 75 mg/mL. The antimicrobial susceptibility test was performed using the disc diffusion method. The results showed an inhibitory effect of crude ethanolic extracts against both Gram-positive and Gram-negative bacteria, also against one fungi. However, crude aqueous extract of leaves has shown no effect against the aforementioned microorganisms. Differently, in vitro St. aureus development has been inhibited by the crude aqueous root extract. In Indian and Chinese folk medicine the parts of the studied plant have been used conventionally. Antimicrobials derived from plants have tremendous therapeutic potential. They effectively treat infectious diseases while simultaneously alleviating some of the adverse effects frequently associated with synthetic alternatives. Taking into consideration of the availability of L. tatarica in Azerbaijani flora and its possible medicinal importance, the results of this investigation open up new horizons for further researches in diverse fields including the pharmaceutical one.