The medium is prepared on a tube by tube basis, using 15 mL glass tubes, stoppered with rubber-lined screw caps.
Add the components in the following order. Once soil and liquid are mixed, the media must be pasteurized within 6 hours. For routine maintenance and transfer, autoclaved media for 15 min, since transfer material typically contains organics and/or bacteria needed for algal growth. In pasteurization, tubes are brought to just over 98 °C, and held at that temperature for 2 hrs. They are then allowed to cool to room temperature for 20-24 hours. This process is repeated for three successive days. It is imperative that media (pasteurized or autoclaved) be allowed to gas-equilibrate for 2 days prior to use.
# | Component | Amount | Stock Solution Concentration | Final Concentration |
---|---|---|---|---|
1 | CR1 Soil | 1 cc | ||
2 | MgCO3 (MCIB CB486) | 0.5 mg/12 mL | 0.5 mM | |
3 | Modified COMBO Medium | 12 mL |