1. Combine the following in 900ml water:
   • 4ml of 250mg/L Resazurin
   • 1ml Trace element solution (see below)
   • 3.5g K₂HPO₄
   • 0.2g Na₂SO₄
   • 5g Yeast extract
   • 1g Tryptone
2. Cook 5-10 min (colour turns first red and then yellow).
3. Cool on ice.
4. Add the following:
   • 10mg biotin
   • 10mg p-aminobenzoic acid
   • 0.5g L-cysteinium chloride (monohydrate)
5. Adjust pH to 7.0.
6. Adjust volume to 1 liter with H₂O
7. Heat slightly to dissolve (do not boil).
8. Distribute into tubes at 5 ml.
9. Autoclave at 121°C for 15 min
10. Add 5 ml sterile 10 g/L glucose

Trace elements solution:

1. Combine the following in 100.0 ml distilled water:
   • MgSO₄ 1.2 g
   • MnSO₄ × 7 H₂O 0.17 g
   • ZnSO₄ × 7 H₂O 29.0 mg
   • CuSO₄ × 5 H₂O 25.0 mg
   • H₂SO₄ 2.8 ml
   • FeCl₃ × 6 H₂O 2.7 g
   • CaCl₂ × 2 H₂O 0.15 g
   • CoCl₂ × 6 H₂O 24.0 mg
   • NaMoO₄ × 2 H₂O 0.24 g
2. Autoclave or filter sterilize.