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Main sol. 120b

Internal Solution-ID: S216

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
K₂HPO₄	0.35	g	0.342	1.962
KH₂PO₄	0.23	g	0.225	1.651
NH₄Cl	0.50	g	0.488	9.128
MgSO₄ x 7 H₂O	0.50	g	0.488	1.981
CaCl₂ x 2 H₂O	0.25	g	0.244	1.661
NaCl	2.25	g	2.197	37.599
FeSO4 x 7 H2O solution (0.1% w/v)	2.00	ml	-	-
Trace element solution SL-10	1.00	ml	-	-
Yeast extract (OXOID)	2.00	g	1.953	-
Casitone (BD BBL)	2.00	g	1.953	-
Na-acetate	2.50	g	2.441	29.761
Sodium resazurin (0.1% w/v)	0.50	ml	4.9e-4	0.002
NaHCO₃	2.50	g	2.441	29.062
2-Mercaptoethanesulfonate (coenzyme M)	0.14	g	0.137	0.961
Methanol (50% v/v)	20.00	ml	15.469	482.765
Wolin's vitamin solution (10x)	1.00	ml	-	-
L-Cysteine HCl x H₂O	0.36	g	0.352	2.002
Na₂S x 9 H₂O	0.36	g	0.352	1.464
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients (except bicarbonate, coenzyme M, vitamins, methanol, cysteine and sulfide) and sparge medium with 80% H₂ and 20% CO₂ gas mixture for 30 - 45 min to make it anoxic. Then add and dissolve bicarbonate, adjust pH to 6.5 and dispense medium under 80% H₂ and 20% CO₂ gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Methanol (50% v/v stock solution) and the reducing agents are each autoclaved separately under 100% N₂ gas atmosphere as concentrated solutions in tightly closed tubes. Vitamins and coenzyme M are prepared under 100% N₂ gas atmosphere and sterilized by filtration. Appropriate volumes of the stock solutions are injected into the sterile medium with hypodermic syringes. Adjust pH of the complete medium to 7.0 - 7.2, if necessary.
2 After inoculation, pressurize culture vessels with sterile 80% H₂ and 20% CO₂ gas mixture to 1 bar overpressure.

Compound

Amount

Unit

Conc. [g/L]

Conc. [mM]

K₂HPO₄

0.35

0.342

1.962

KH₂PO₄

0.23

0.225

1.651

NH₄Cl

0.50

0.488

9.128

MgSO₄ x 7 H₂O

0.50

0.488

1.981

CaCl₂ x 2 H₂O

0.25

0.244

1.661

NaCl

2.25

2.197

37.599

FeSO4 x 7 H2O solution (0.1% w/v)

2.00

Trace element solution SL-10

1.00

Yeast extract
(OXOID)

2.00

1.953

Casitone
(BD BBL)

2.00

1.953

Na-acetate

2.50

2.441

29.761

Sodium resazurin
(0.1% w/v)

0.50

4.9e-4

0.002

NaHCO₃

2.50

2.441

29.062

2-Mercaptoethanesulfonate
(coenzyme M)

0.14

0.137

0.961

Methanol
(50% v/v)

20.00

15.469

482.765

Wolin's vitamin solution (10x)

1.00

L-Cysteine HCl x H₂O

0.36

0.352

2.002

Na₂S x 9 H₂O

0.36

0.352

1.464

Distilled water

1000.00

1 Dissolve ingredients (except bicarbonate, coenzyme M, vitamins, methanol, cysteine and sulfide) and sparge medium with 80% H₂ and 20% CO₂ gas mixture for 30 - 45 min to make it anoxic. Then add and dissolve bicarbonate, adjust pH to 6.5 and dispense medium under 80% H₂ and 20% CO₂ gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Methanol (50% v/v stock solution) and the reducing agents are each autoclaved separately under 100% N₂ gas atmosphere as concentrated solutions in tightly closed tubes. Vitamins and coenzyme M are prepared under 100% N₂ gas atmosphere and sterilized by filtration. Appropriate volumes of the stock solutions are injected into the sterile medium with hypodermic syringes. Adjust pH of the complete medium to 7.0 - 7.2, if necessary.

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

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Main sol. 120b

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Main sol. 120b

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