Medium data

Search for medium no. = [431].

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

431 BN3 MEDIUM

Basal medium (see below)	`1.0`	L
Trace element solution (see below)	`1.0`	ml
Selenite-tungstate solution (see below)	`1.0`	ml
Trace vitamins (see Medium No. 197)	`10.0`	ml
Vitamin B₁₂	`5.0`	µg
Glucose	`3.6`	g
Yeast extract (BD-Difco)	`0.5`	g
Tryptone (BD-Difco)	`1.5`	g
Sulfur (powder)	`20.0`	g
NaHCO₃	`3.4`	g
Resazurin	`0.5`	mg
Na₂S·9H₂O	`0.36`	g

Autoclave Basal medium supplemented with Trace element solution, Selenite-tungstate solution and resazurin under N₂-CO₂ (4:1, v/v). Separately autoclave glucose, yeast extract and tryptone (10% each) and add to the medium. Filter-sterilize Trace vitamins and NaHCO₃ (8%) solution and add to the medium. Steam sulfur for 3 hr on each of 3 successive days. Distribute the medium into culture vessels containing sulfur under a N₂-CO₂ (4:1, v/v) gas mixture and seal with butyl rubber stoppers. Before inoculation, reduce the medium with 5% Na₂S·9H₂O solution (autoclaved under a N₂ atmosphere). Readjust pH to 7.2, if necessary.
Basal medium:

NaCl	`1.0`	g
MgCl₂·6H₂O	`0.4`	g
CaCl₂·2H₂O	`0.075`	g
NH₄Cl	`0.25`	g
KH₂PO₄	`0.2`	g
KCl	`0.5`	g
Distilled water	`1.0`	L

Trace element solution:

Distilled water	`987.0`	ml
25% HCl	`12.5`	ml
FeSO₄·7H₂O	`2.1`	g
H₃BO₃	`30.0`	mg
MnCl₂·4H₂O	`100.0`	mg
CoCl₂·6H₂O	`190.0`	mg
NiCl₂·6H₂O	`24.0`	mg
CuCl₂·2H₂O	`2.0`	mg
ZnSO₄·7H₂O	`144.0`	mg
Na₂MoO₄·2H₂O	`36.0`	mg

Selenite-tungstate solution:

NaOH	`0.4`	g
Na₂SeO₃·5H₂O	`6.0`	mg
Na₂WO₄·2H₂O	`8.0`	mg
Distilled water	`1.0`	L