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Main sol. 897

Internal Solution-ID: S1847

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
NaCl	13.65	g	13.542	231.719
MgSO₄ x 7 H₂O	3.50	g	3.472	14.088
MgCl₂ x 6 H₂O	2.75	g	2.728	13.419
KH₂PO₄	0.50	g	0.496	3.645
(NH₄)₂SO₄	0.25	g	0.248	1.877
CaCl₂ x 2 H₂O	0.38	g	0.377	2.564
KCl	0.33	g	0.327	4.391
NaBr	0.05	g	0.05	0.482
H₃BO₃	15.00	mg	0.015	0.241
SrCl2 x 6 H₂O (0.1% w/v)	7.00	ml	0.007	0.026
KI (0.01% w/v)	0.50	ml	5.0e-5	1.9e-4
Sodium resazurin (0.1% w/v)	0.50	ml	5.0e-4	0.002
Sulfur (powdered)	5.00	g	4.96	154.672
Meat extract	1.00	g	0.992	-
Na₂S x 9 H₂O	0.20	g	0.198	0.826
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients except sulfur, meat extract and sulfide, adjust pH to 5.0 - 5.5 with 2 N H₂SO₄ and sparge medium with 80% H₂ and 20% CO₂ gas mixture for 30 - 45 min to make it anoxic. Distribute medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials already containing the appropriate amount of sulfur, only to 30% of their volume to allow for a large head space. For sterilization heat cultivation vessels in boiling water for 1 -2 hours on each of 3 successive days. Add meat extract and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas atmosphere.
2 After inoculation pressurize the vessels with sterile 80% H₂ and 20% CO₂ gas mixture to 2 bar overpressure.

Compound

Amount

Unit

Conc. [g/L]

Conc. [mM]

NaCl

13.65

13.542

231.719

MgSO₄ x 7 H₂O

3.50

3.472

14.088

MgCl₂ x 6 H₂O

2.75

2.728

13.419

KH₂PO₄

0.50

0.496

3.645

(NH₄)₂SO₄

0.25

0.248

1.877

CaCl₂ x 2 H₂O

0.38

0.377

2.564

KCl

0.33

0.327

4.391

NaBr

0.05

0.482

H₃BO₃

15.00

0.015

0.241

SrCl2 x 6 H₂O
(0.1% w/v)

7.00

0.007

0.026

KI
(0.01% w/v)

0.50

5.0e-5

1.9e-4

Sodium resazurin
(0.1% w/v)

0.50

5.0e-4

0.002

Sulfur
(powdered)

5.00

4.96

154.672

Meat extract

1.00

0.992

Na₂S x 9 H₂O

0.20

0.198

0.826

Distilled water

1000.00

1 Dissolve ingredients except sulfur, meat extract and sulfide, adjust pH to 5.0 - 5.5 with 2 N H₂SO₄ and sparge medium with 80% H₂ and 20% CO₂ gas mixture for 30 - 45 min to make it anoxic. Distribute medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials already containing the appropriate amount of sulfur, only to 30% of their volume to allow for a large head space. For sterilization heat cultivation vessels in boiling water for 1 -2 hours on each of 3 successive days. Add meat extract and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas atmosphere.

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

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Main sol. 897

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Main sol. 897

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