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This study was carried out to develop the best medium of new oyster mushroom for bottle culture. The new oyster mushroom cv. Miso is a Pleurotus ostreatus developed at the Chungnam Agricultural Research & Extension Services. For the bottle culture media, poplar sawdust+beet pulp+cottonseed meal (5:3:2), poplar sawdust+wheat husk meal (8:2), poplar sawdust+beet pulp+cottonseed meal (6:2:2), poplar sawdust+beet pulp+cottonseed meal (4:4:2), and poplar sawdust+beet pulp+wheat husk meal (7:1:2) were used in 850cc PP bottle. The pH was 5~6 and the C/N ratio 19.7~28.3 in bottle culture media. The time of pinhead formation was 5 to 6 days. For the fruiting body formation after inoculation took 29~31 days. The yield of fruiting body of poplar sawdust+beet pulp+wheat husk meal (7:1:2) medium was the highest at 110.4g/bottle compared to other media. Therefore, such cultivation medium would be appropriate for the commercial production of bottle culture in the new oyster mushroom ‘Miso’.