Medium data

Search for medium no. = [513].

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

513 CALDOCOCCUS MEDIUM

NaCl	`0.1`	g
CaCl₂·2H₂O	`0.1`	g
FeSO₄·7H₂O	`0.05`	g
Yeast extract (BD-Difco)	`0.1`	g
Casamino acids (BD-Difco)	`0.1`	g
Peptone	`0.8`	g
Trace vitamins (see Medium No. 197)	`10.0`	ml
Trace mineral solution (see below)	`1.0`	ml
Sulfur	`2.0`	g
Resazurin	`1.0`	mg
Na₂S·9H₂O	`0.5`	g
Distilled water	`1.0`	L

Mix ingredients except vitamins, sulfur and Na₂S·9H₂O, adjust pH to 2.7 with H₂SO₄ and autoclave. Separately autoclave 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. Distribute the medium into culture vessels (e.g., 20 ml per 100 ml serum bottles) containing Trace vitamins and sulfur, replace the gas phase with a H₂-CO₂ (80:20) gas atmosphere and seal with butyl rubber stoppers. Prior to inoculation, add sterile Na₂S·9H₂O solution and adjust pH to 3.0 with sterile 1 N H₂SO₄. Pressurize the inoculated bottle to 200 kPa H₂-CO₂ (80:20).
Trace mineral solution:

MnSO₄·xH₂O	`5.0`	g
CoSO₄7H₂O	`1.0`	g
ZnSO₄·7H₂O	`1.0`	g
CuSO₄·5H₂O	`0.1`	g
H₃BO₃	`0.1`	g
Na₂MoO₄·2H₂O	`0.1`	g
Distilled water	`1.0`	L