429. RICH ORGANIC MEDIUM WITH 2% NaCl (ROMNa)

NaCl 20.0 g

MgSO₄ × 7 H₂O 0.5g

KH₂PO₄ 0.3 g

NH₄Cl 0.3 g

KCl 0.3 g

CaCl₂ × 6 H₂O 0.05 g

Acetate Na 1.0 g

Yeast extract 1.0 g

Bactopeptone 0.5 g

Casamino acids 0.5 g

Trace element solution (see below) 2 ml

Vitamin solution (see below) 2 ml

Distilled water 1000.0 ml

Adjust pH to 5.9 and autoclave. Adjust pH to 6.8-7.0 with 0.5N NaOH

Trace element solution:

FeSO₄ × 7 H₂O 0.3 g

MnCl₂ × 4 H₂O 0.003 g

CoCl₂ × 6 H₂O 0.005 g

CuCl₂ × 5 H₂O 0.001 g

NiCl₂ × 6 H₂O 0.002 g

Na₂MoO₄ × 2 H₂O 0.003 g

ZnSO₄ × 7 H₂O 0.005 g

H₃BO₃ 0.002 g

Dissolve in 0.5 l of distilled water and adjust pH to 3.0. Fill to 1 l and refrigerate

Vitamin solution:

Nicotinic acid 0.4 g

Thiamin 0.4 g

Biotin 0.08 g

Vitamin B₁₂ 0.02 g

Filter sterilize and refrigerate