Medium data

Search for medium no. = [308].

Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

308 ACIDILOBUS MEDIUM

NH₄Cl	`0.33`	g
KH₂PO₄	`0.33`	g
KCl	`0.33`	g
CaCl₂·2H₂O	`0.33`	g
MgCl₂·6H₂O	`0.33`	g
FeCl₂ solution (see Medium No. 187)	`1.0`	ml
Trace element solution (see Medium No. 187)	`1.0`	ml
Trace vitamins (see Medium No. 197)	`10.0`	ml
Yeast extract (BD-Difco)	`0.3`	g
Sulfur (powder)	`10.0`	g
Na₂S·9H₂O	`0.5`	g
Resazurin	`1.0`	mg
Distilled water	`1.0`	L

Mix ingredients, except yeast extract, vitamins, sulfur and Na₂S·9H₂O, and autoclave under a N₂-CO₂ (80:20) gas atmosphere. Separately autoclave 10% (w/v) yeast extract solution and 5% (w/v) Na₂S·9H₂O solution under a N₂ atmosphere. Steam sulfur for 3 hr on each of 3 successive days. Filter-sterilize Trace vitamins solution while gassing with N₂. Aseptically and anaerobically add the yeast extract and vitamin solutions, and sulfur to the medium under a N₂-CO₂ (80:20) gas atmosphere. Prior to inoculation, add sterile Na₂S·9H₂O solution, and adjust pH to 3.5 - 3.8 with sterile 1 N H₂SO₄. Pressurize the inoculated bottle to 100 kPa N₂-CO₂ (80:20).