The plant Rose has many varieties that Rosa foetida Hermm (Persian yellow rose) is a member of Rosacea tribe. This flower grows in some countries, including North West, West and Center of Iran. In this study, the plant was purchased and named by herbarium department of Shaheed Beheshti University of medical sciences. Its essential oil was obtained byhydro distillation and analyzed by GC and GC/MS. The major components of its essence were Thymol (28.7%), NONADECANE (14.2%), 1,8-Anydro-cis-alpha-copaene 8-01 (13.3%), Heneicosane (11.9%), 1-NONADECANE(7.3%), Carvacrol (5.6%). The whole extract deleted by succylate apparatus and methanol as solvent and concentrated in rotary distillatory. Antibacterial activity of methanol extract was studied on 5 major gram positive and gram negative bacteria by both disk diffusion and cup plate methods. MIC was determined based on microdilution protocol.The inhibition zone for 0.75g/cc of whole extract against S.aureus, S.typhi, E.coli and P.aeruginosa were 19, 14, 12, 25 mm, respectably. The later diameter (25mm) was equal to gentamicin as positive control. MIC for P.aeroginosa, S.aureus, E.coli, S.typhi was 0.008g/cc, 0.015g/cc and 0.03 g/cc, respectably. In conclusion, comparison of both disk diffusion and cup plate method showed better result by cup plate (P<0.05). Further studies on toxicity, improved formula and in-vivo tests are recommended.