239. METHYLOPHAGA ALCALICA MEDIUM

KH₂PO₄ 1.0 g

KNO₃ 1.0 g

MgSO₄ × 7 H₂O 0.22 g

NaCl 30.0 g

Na₂CO₃ 5.0 g

Trace element solution (see below) 1.0 ml

Vitamin B₁₂ 0.02 mg

Methanol 10.0 ml

Distilled water 1000.0 ml

Final pH 9.5

Trace elements solution:

Ferric citrate 30.0 mg

CaCl₂ × 2 H₂O 30.0 mg

MgCl₂ × 4 H₂O 5.0 mg

ZnSO₄ × 7 H₂O 5.0 mg

CuSO₄ × 5 H₂O 0.5 g

Distilled water 1000.0 ml

Prepare the medium without the Na₂CO₃, vitamin B₁₂ and methanol. Basal medium, trace element and Na₂CO₃ autoclave separately at 121°C 15 min. Add the vitamin B₁₂ solution from a filter-sterilized stock solution and filter-sterilized methanol to the cooled medium. When preparing liquid media cool the mineral salts solution and Na₂CO₃ to room temperature before mixing. When preparing agar add 2.0 % agar to the mineral salt solution and autoclave. Cool the Na₂CO₃ stock solution and agar to 50-55°C before mixing.

 

Updated 1.02.2024