Studies for the discovery of microorganisms with different metabolic properties in extreme environments are increasing every year. Within the scope of this study, samples were taken from, an extreme environment, İnsuyu cave. Antibiotic resistance in the obtained isolates was investigated by cultural and molecular biology methods. As a result of Minimum Inhibition Concentration (MIC) test, 64 cefpodoxim, 6 rifampicin and gentamycin, 18 vancomycin, 15 ampicillin, 44 clindamycin, 48 penicillin resistant strains were detected. Pseudomonas spp. (2G-2), Pseudomonas jesseni (2J), Sphingopyxis fribergensis (4D), Microbacterium yannicii (4M), Flavobacterium chungangense (6B), Rhodococcus spp. (7A), Flavobacterium resistens (9D), Pseudomonas spp. (K-4F), Pseudomonas spp. (K-4G), Buttiauxella agrestis (K-15A) were found to have multidrug resistant by cultural methods. These bacteria and susceptible strains, Flavobacterium chungangense (K-1E), Rhodococcus erythropolis (K-11G) and Pseudomonas spp. (K-15G), as negative controls were selected for the identification of antibiotic resistance genes by PCR based methods. Eight different gene regions, aminoglycoside 2''-O-nucleotyltransferase (aadB), beta lactamase (blaCTXM3, blaSHV, blaTEM), aminoglycoside resistance protein (strA), rifampin ADP- ribosyltransferase (arr2- int2a), vancomycin (vanC) have been detected in Pseudomonas jesseni (2J) and Pseudomonas spp. (K-4G).