298. METHYLOROSULA POLARIS MEDIUM

Mineral salts solution:

(NH₄)₂SO₄ 0.05 g

MgCl₂ × 6 H₂O 0.04 g

KH₂PO₄ 0.07 g

Trace element solution (see below) 1.0 ml

Distilled water 1000.0 ml

Trace element solution:

Na₂EDTA 500.0 mg

FeSO₄ × 7 H₂O 200.0 mg

ZnSO₄ × 7 H₂O 10.0 mg

MgCl₂ × 6 H₂O 3.0 mg

H₃BO₃ 30.0 mg

CoCl₂ × 6 H₂O 20.0 mg

CaCl₂ × 2 H₂O 1.0 mg

NiCl₂ × 6 H₂O 2.0 mg

Na₂MoO₄ × 2 H₂O 3.0 mg

Distilled water 1000.0 ml

Prepare the liquid medium. Adjust to pH 6.0. Dispense the medium in tubes or flasks fitted with screw caps. Sterilize by autoclaving at 121°C for 15 min. Before inoculation add filter sterilized CH₃OH to a final concentration of 0.5% v/v.