image-logoThe Read Lab

Making Procyclic Form Media

  1. Find the 10 L plastic bucket.
  2. Rinse bucket and 2 L cylinder with distilled water and then with MilliQ water.
  3. Pour 8 L of MilliQ water into the bucket.
  4. Wash the big stir bar with MilliQ.
  5. Stir the water.
  6. Add 1/2 a bottle of HyClone SM media powder (296 g). Measure 2x 148 g.
  7. Stir for at least 1/2 hour.
  8. Meanwhile, thaw MEM vitamins and prepare fresh NaOH (20g of pellets -> 50 mL - use MilliQ.
  9. Add 25 mL MEM vitamins.
  10. Adjust pH to 7.5 with fresh NaOH (pour in about 25 mL and then go dropwise).
  11. Increase the volume to approx. 9.4L.
  12. Stir for 1/2 hour or longer.
  13. Filter sterilize (low protein binding filter) - 900 mL/bottle. Use the graduation on bottle to gauge the right amount.

Download Protocol: Procyclic Form Media Protocol
(note requires Microsoft Word to open)