284. METHYLOCYSTIS MEDIUM

KNO₃ 0.2 g

KH₂PO₄ 0.1 g

MgSO₄ × 7 H₂O 0.05 g

CaCl₂ × 2 H₂O 0.01 g

NaCl 0.01 g

Trace element solution (see below) 1.0 ml

Distilled Water 1000.0 ml

Final pH 5.8-6.2.

Trace elements solution:

EDTA 5.0 g

CuCl₂ × 2 H₂O 0.1 g

FeSO₄ × 7 H₂O 2.0 g

ZnSO₄ × 7 H₂O 0.1 g

NiCl₂ × 6 H₂O 0.02 g

CoCl₂ × 6 H₂O 0.2 g

Na₂MoO₄ 0.03 g

Distilled water 1000.0 ml

Dispense the medium into serum bottles with a medium to headspace ratio of about 1:4. Sterilize by autoclaving at 121°C for 15 min. Add 20% methane to the gas phase. Incubate the cultures with shaking.