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 processed (pasteurized or autoclaved) within 6 hrs. 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 hrs. 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|
|4||Pasteurized Seawater||1 mL|