ATCC medium: 132 Zymobacterium medium
Tryptone (BD 211705)........20.0 g
Orotic acid..................2.0 g
NaOH.........................0.5 g
Na3PO4 . 12H2O................1.5 g
Sodium thioglycollate........1.0 g
Riboflavin...................0.015 g
Agar (if needed)............15.0 g
Distilled water..............1.0 L
Add ingredients in the order listed at room temperature. Orotic acid will not dissolve completely. The final pH should be 7.9. If it is found to be higher, add a small amount of NaH2PO4. Agitate the mixture each time prior to dispensing into test tubes. Autoclave at 121C for 15- 20 minutes. If necessary, agitate the tubes slightly after autoclaving to dissolve the remaining orotic acid.