Composition per 1070.0mL:

Solution A…… 1.0L

Solution B …… 60.0mL

Solution C …… 10.0mL

pH 7.2 ± 0.2 at 25°C

Solution A:

Composition per liter:

NaCl ……25.0g

Agar ……15.0g

MgSO₄·7H₂O……5.0g

Casamino acids ……2.5g

l-Carrageenan……2.5g

NaNO₃……2.0g

CaCl₂·2H₂O……0.2g

KCl……0.1g

Preparation of Solution A: Add components to distilled/deionized water and bring volume to 1.0L.

Mix thoroughly. Gently heat and bring to boiling.

Autoclave for 15 min at 15 psi pressure –121°C.

Solution B:

Composition per 100.0mL:

Na₂HPO₄·2H₂O……3.56g

Preparation of Solution B: Add component to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure –121°C.

Solution C:

Composition per 100.0mL:

FeSO₄·7H₂O……0.3g

Preparation of Solution C: Add component to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure –121°C.

Preparation of Medium: Aseptically add

60.0mL of sterile solution B and 10.0mL of sterile solution C to 1.0L of sterile solution A. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation and maintenance of ATCC strain 43554.