This study evaluates the sensitivity of various screening methods for determining the antibacterial activity of natural products. Fourteen natural products from plants, fungi, and lichens were tested against *Staphylococcus aureus* and *Escherichia coli* using two agar diffusion methods (well and disc variants), two bioautographic methods (direct and indirect variants), and a microdilution assay. The results showed that the well-variant of the agar diffusion method was more sensitive than the disc-variant, while the direct-variant of the bioautographic method exhibited greater sensitivity compared to the indirect-variant. Both bioautographic and diffusion techniques demonstrated similar sensitivity, but the latter provided more suitable conditions for microbial growth. The study also discusses the optimal conditions for determining the minimal inhibitory concentration (MIC) of natural products. The findings suggest that the well-variant of the agar diffusion method and the direct-variant of the bioautographic method are more effective for screening natural products with antibacterial activity.This study evaluates the sensitivity of various screening methods for determining the antibacterial activity of natural products. Fourteen natural products from plants, fungi, and lichens were tested against *Staphylococcus aureus* and *Escherichia coli* using two agar diffusion methods (well and disc variants), two bioautographic methods (direct and indirect variants), and a microdilution assay. The results showed that the well-variant of the agar diffusion method was more sensitive than the disc-variant, while the direct-variant of the bioautographic method exhibited greater sensitivity compared to the indirect-variant. Both bioautographic and diffusion techniques demonstrated similar sensitivity, but the latter provided more suitable conditions for microbial growth. The study also discusses the optimal conditions for determining the minimal inhibitory concentration (MIC) of natural products. The findings suggest that the well-variant of the agar diffusion method and the direct-variant of the bioautographic method are more effective for screening natural products with antibacterial activity.