Tissue culture is the most commonly used method for producing virus-free potato seeds. Contamination is one of the most frequent problems that occur during the tissue culture, that causes microbial infiltration in the air, and is a fatal factor that can result in enormous losses, especially in large-scale cultivation for industrial production. Therefore, it is important to know what kinds of fungi and bacteria are involved in the contamination of potato tissue culture, and to find suitable fungicides and antibiotics to inhibit microbial contamination without affecting the growth of potato tissue culture. In this study, we examined six fungicides such as pyraclostrobin, tebuconazole, carbendazim, mancozeb, chlorothalonil, fluazinam, and five antibiotics such as streptomycin, ampicillin, chloramphenicol, vancomycin and penicillin G for controlling microbial contamination. In the experiment with the fungicides and antibiotics supplemented plants, the treatments with less significant difference from the untreated plants were selected, and the concentrations of the agents were selected by lowering the concentration of the chemicals. Three fungicides such as pyraclostrobin, chlorothalonil, fluazinam and Three antibiotics such as ampicillin, vancomycin, and penicillin G did not inhibit plant growth even at a concentration of 10 ppm. In this study, four fungi such as Aspergillus niger, Cladosporium sphaerosperum, Simpicillium lamellicola and Purpureocillium lilacinum and five species of bacteria such as Achromobacter sp., Stenotrophomonas sp., Ochrobacterum sp., Methylobacterium sp., Staphylococcus sp. were identified, which were contaminated of the media. After identifying the fungi and bacteria that cause contamination of the media, they were treated with medicines to select the medicines and concentrations that inhibited the growth of the contaminants. Among the fungi, fluazinam showed the highest rate of mycelial growth inhibition against A. niger, and pyraclostrobin showed the highest mycelial growth inhibition rate on C. sphaerosperum, S. lamellicola and P. lilacinum. Staphylococcus sp. was inhibited by 25 ppm of vancomycin and Stenotrophomonas sp. was inhibited by penicillin G of 25 ppm. Based on the experimental results, the list of the drugs that inhibit the growth of plants and the inhibitors of the growth of contaminants were selected and concentrations were determined. As a result, 200ppm of fungicide and 25ppm of antibiotics were added to the medium. In the final selection, the antibiotics were vancomycin and penicillin G and the fungicides were pyraclostrobin and fluazinam. As a result of the addition of two drugs to the medium, the vancomycin 25ppm + pyraclostrobin 200ppm and penicillin G 25ppm + fluazinam 200ppm did not inhibit the growth of the plant and the medium contamination did not occur. Therefore, it was concluded that tissue culture using MS medium containing 25 ppm of vancomycin, 200 ppm of pyraclostrobin, 25 ppm of penicillin G, and 200 ppm of fluazinam in potato tissue culture were not inhibit growth of tissue culture seedlings with high inhibitory effect against contaminants.