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Main sol. 883

Internal Solution-ID: S1829

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Trisodium citrate x 2 H₂O	2.94	g	2.908	9.888
(NH₄)₂SO₄	1.30	g	1.286	9.731
KH₂PO₄	0.28	g	0.277	2.035
MgSO₄ x 7 H₂O	0.25	g	0.247	1.003
CaCl₂ x 2 H₂O	0.07	g	0.069	0.471
FeCl₃ x 6 H₂O	0.02	g	0.02	0.073
Allen's trace element solution	10.00	ml	-	-
Sodium resazurin (0.1% w/v)	0.50	ml	4.9e-4	0.002
Sulfur (powdered)	10.00	g	9.891	308.425
Yeast extract (OXOID)	0.50	g	0.495	-
Wolin's vitamin solution (10x)	1.00	ml	-	-
Na₂S x 9 H₂O	0.50	g	0.495	2.059
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients except sulfur, yeast extract, vitamins and sulfide, then adjust the pH to 3.5 with 4 N H₂SO₄. Sparge medium with 100% N₂ gas for 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials which contain already the appropriate amount of sulfur, only to 30% of their volume to allow for a large headspace. Autoclave at 105°C for 20 min. Prior to inoculation add yeast extract, vitamins (sterilized by filtration) and sulfide from anoxic stock solutions prepared under 100% N₂ gas atmosphere. The final pH of the complete medium should be around 4.0.
2 After inoculation, pressurize the cultivation vessels to 1 bar overpressure with sterile 80% H₂ and 20% CO₂ gas mixture.

Compound

Amount

Unit

Conc. [g/L]

Conc. [mM]

Trisodium citrate x 2 H₂O

2.94

2.908

9.888

(NH₄)₂SO₄

1.30

1.286

9.731

KH₂PO₄

0.28

0.277

2.035

MgSO₄ x 7 H₂O

0.25

0.247

1.003

CaCl₂ x 2 H₂O

0.07

0.069

0.471

FeCl₃ x 6 H₂O

0.02

0.073

Allen's trace element solution

10.00

Sodium resazurin
(0.1% w/v)

0.50

4.9e-4

0.002

Sulfur
(powdered)

10.00

9.891

308.425

Yeast extract
(OXOID)

0.50

0.495

Wolin's vitamin solution (10x)

1.00

Na₂S x 9 H₂O

0.50

0.495

2.059

Distilled water

1000.00

1 Dissolve ingredients except sulfur, yeast extract, vitamins and sulfide, then adjust the pH to 3.5 with 4 N H₂SO₄. Sparge medium with 100% N₂ gas for 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials which contain already the appropriate amount of sulfur, only to 30% of their volume to allow for a large headspace. Autoclave at 105°C for 20 min. Prior to inoculation add yeast extract, vitamins (sterilized by filtration) and sulfide from anoxic stock solutions prepared under 100% N₂ gas atmosphere. The final pH of the complete medium should be around 4.0.

2 After inoculation, pressurize the cultivation vessels to 1 bar overpressure with sterile 80% H₂ and 20% CO₂ gas mixture.

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Main sol. 883

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