Turkey tail mushroom (Trametes versicolor) is a medicinal mushroom used as a health-promoting pharmaceutical, extending life, and adjuvant therapy for cancer patients. The mycelia and fruiting bodies of the Turkey tail mushroom contain various biologically active compounds such as polysaccharides, glucans, phenolics, flavonols, coumarins, isoflavonoids, triterpenoids, and sterols. This study was carried out to choose the optimal culture conditions, including culture pH, temperature, medium, and substrate composition, for the growth of the mycelia and fruiting bodies of the T. versicolor mushroom Tra-6 strain, collected in Lao Cai, Vietnam. The experimental factors were examined individually in a randomized complete block design. Research results show that the mycelia of the Turkey tail mushroom Tra-6 strain quickly grew on MCM and YEA media at pH 8 and 25 ± 1C. Sucrose, yeast extract, MgSO4.7H2O, KCl, and KH2PO4 were the optimal carbon, nitrogen, and mineral salt sources for this strain, respectively. Tra-6 strain had the ability to form fruiting bodies when growing on substrate from agricultural by-products. The highest yield of 44.4kg dry mushroom per ton of dry substrate was obtained on a substrate consisting of 55% sawdust, 24% crushed corncobs, 20% rice bran, and 1% CaCO3 powder.