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Main sol. 1360

Internal Solution-ID: S2740

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
NaCl	26.33	g	26.044	445.645
MgCl₂ x 6 H₂O	1.50	g	1.484	7.298
KCl	0.45	g	0.445	5.971
(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	-	-
Na-acetate	1.25	g	1.236	15.072
Trisodium citrate x 2 H₂O	2.94	g	2.908	9.888
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 (DIFCO)	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, vitamins, yeast extract and sulfide), adjust pH to 3.5 with 10 N H₂SO₄ and sparge medium with 80% N₂ and 20% CO₂ gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into serum vials (e.g., 20 ml medium in 100 ml bottles) that contain already the appropriate amount of sulfur. Sterilize medium by heating cultivation vessels in a water bath to 90 - 100°C for 1 - 2 hours on each of 3 successive days. Add vitamins (sterilized by filtration) and yeast extract from sterile anoxic stock solutions prepared under 100% N₂ gas. Prior to inoculation, add sulfide from a sterile anoxic stock solution prepared under 100% N₂ gas and adjust pH of complete medium to 4.5, if necessary.

Compound

Amount

Unit

Conc. [g/L]

Conc. [mM]

NaCl

26.33

26.044

445.645

MgCl₂ x 6 H₂O

1.50

1.484

7.298

KCl

0.45

0.445

5.971

(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

Na-acetate

1.25

1.236

15.072

Trisodium citrate x 2 H₂O

2.94

2.908

9.888

Sodium resazurin
(0.1% w/v)

0.50

4.9e-4

0.002

Sulfur , powdered

10.00

9.891

308.425

Yeast extract
(DIFCO)

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, vitamins, yeast extract and sulfide), adjust pH to 3.5 with 10 N H₂SO₄ and sparge medium with 80% N₂ and 20% CO₂ gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into serum vials (e.g., 20 ml medium in 100 ml bottles) that contain already the appropriate amount of sulfur. Sterilize medium by heating cultivation vessels in a water bath to 90 - 100°C for 1 - 2 hours on each of 3 successive days. Add vitamins (sterilized by filtration) and yeast extract from sterile anoxic stock solutions prepared under 100% N₂ gas. Prior to inoculation, add sulfide from a sterile anoxic stock solution prepared under 100% N₂ gas and adjust pH of complete medium to 4.5, if necessary.

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Main sol. 1360

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