Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended Medium Renewal: Twice per week Remove medium, add fresh 0.25% trypsin solution for 2 to 3 minutes, remove trypsin and let the culture sit at 37C for 5 to 10 minutes. Add fresh medium, aspirate and dispense into new flasks. Subculture every 6 to 8 days. |