Mix ingregients except vitamin solution, adjust pH to 6.5 and autoclave under a N₂ atmosphere. Aspectically and anaerobically add filter-sterile vitamin solution to the medium. Final pH is 6.5.

*Trace element solution

First dissolve NTA and adjust pH to 6.5 with NaOH, and then add minerals. Final pH is 7.0.

**Vitamin solution

Adjust pH to 7.0 with NaOH.