|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.