371. MINERAL MEDIUM “P” FOR METHANOTROPHIC BACTERIA

Na₂HPO₄ × 12 H₂O 1.5 g

KH₂PO₄ 0.7 g

KNO₃ 1.0 g

MgSO₄ × 7 H₂O 0.2 g

CaCl₂ × 2 H₂O 0.02 g

NaCl 30.0 g

NaHCO₃ 4.2 g

Na₂CO₃ 0.05 g

Trace element solution (see below) 1.0 ml

Distilled water 1000.0 ml

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

Prepare the medium without the NaHCO₃ and Na₂CO₃. Basal medium, trace element solution, NaHCO₃ and Na₂CO₃ sterilize separately at 121°C for 15 min. When preparing liquid media cool the mineral salts solution, NaHCO₃ and Na₂CO₃ to room temperature before mixing. When preparing agar add 2.0 % agar to the mineral salt solution and autoclave. Cool the NaHCO₃ and Na₂CO₃ stock solution and agar to 50-55°C before mixing.

The pH of the complete medium should be at 9.0.

 

Updated 31/05/2020