341. RUTHENIBACTERIUM MEDIUM

Trypticase peptone 5.0 g

Peptone 5.0 g

Yeast extract 10.0 g

Beef extract 5.0 g

Glucose 5.0 g

K₂HPO₄ 2.0 g

Tween 80 1.0 ml

L-Cysteine-HCl × H₂O 0.5 g

Resazurin 1.0 mg

Salt solution (see below) 40.0 ml

Distilled water 950.0 ml

Haemin solution (see below) 10.0 ml

Vitamin K₁ solution (see below) 0.2 ml

The vitamin K₁, haemin solution and the cysteine are added after the medium has been boiled and cooled under CO₂. Adjust pH to 7.2 using 8 N NaOH. Distribute under N₂ and autoclave.

Salt solution:

CaCl₂ × 2 H₂O 0.25 g

MgSO₄ × 7 H₂O 0.5 g

K₂HPO₄ 1.0 g

KH₂PO₄ 1.0 g

NaHCO₃ 10.0 g

NaCl 2.0 g

Distilled water 1000.0 ml

Haemin solution: Dissolve 50 mg haemin in 1.0 ml 1 N NaOH; make up to 100.0 ml with distilled water. Store refrigerated.

Vitamin K₁ solution: Dissolve 0.1 ml of vitamin K₁ in 20.0 ml 95% ethanol and filter sterilize. Store refrigerated in a brown bottle.

 

Updated 01.02.2024