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Main sol. 829c

Internal Solution-ID: S1683

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Sea Salt (SIGMA)	30.00	g	29.674	-
NH₄Cl	1.00	g	0.989	18.492
KH₂PO₄	0.35	g	0.346	2.544
MES [2-(N-morpholino) ethane sulfonic acid]	1.95	g	1.929	9.879
Modified Wolin's mineral 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
Na₂CO₃	0.50	g	0.495	4.666
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, carbonate, vitamins and sulfide), boil medium for 1 min, then cool to room temperature under 80% H₂ and 20% CO₂ gas atmosphere. Adjust pH to 6.0 and dispense under 80% H₂ and 20% CO₂ 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 head space. Autoclave at a temperature of 110°C for 20 min! Add vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas atmosphere and carbonate from a sterile anoxic stock solution prepared under 80% N₂ and 20% CO₂ gas mixture. Vitamins are sterilized by filtration. Adjust pH of the complete medium to 6.0 - 6.5, if necessary.
2 After inoculation use 1 bar overpressure of sterile 80% H₂ and 20% CO₂ gas mixture.

Compound

Amount

Unit

Conc. [g/L]

Conc. [mM]

Sea Salt
(SIGMA)

30.00

29.674

NH₄Cl

1.00

0.989

18.492

KH₂PO₄

0.35

0.346

2.544

MES [2-(N-morpholino) ethane sulfonic acid]

1.95

1.929

9.879

Modified Wolin's mineral solution

10.00

Sodium resazurin
(0.1% w/v)

0.50

4.9e-4

0.002

Sulfur , powdered

10.00

9.891

308.425

Na₂CO₃

0.50

0.495

4.666

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, carbonate, vitamins and sulfide), boil medium for 1 min, then cool to room temperature under 80% H₂ and 20% CO₂ gas atmosphere. Adjust pH to 6.0 and dispense under 80% H₂ and 20% CO₂ 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 head space. Autoclave at a temperature of 110°C for 20 min! Add vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas atmosphere and carbonate from a sterile anoxic stock solution prepared under 80% N₂ and 20% CO₂ gas mixture. Vitamins are sterilized by filtration. Adjust pH of the complete medium to 6.0 - 6.5, if necessary.

2 After inoculation use 1 bar overpressure of sterile 80% H₂ and 20% CO₂ gas mixture.

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Main sol. 829c

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