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Main sol. 684

Internal Solution-ID: S1432

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Na₂HPO₄ x 2 H₂O	0.53	g	0.528	3.722
KH₂PO₄	0.41	g	0.409	3.004
NH₄Cl	0.30	g	0.299	5.592
CaCl₂ x 2 H₂O	0.11	g	0.11	0.746
MgCl₂ x 6 H₂O	0.10	g	0.1	0.49
NaCl	0.30	g	0.299	5.118
Trace element solution SL-10	1.00	ml	-	-
Selenite-tungstate solution	1.00	ml	-	-
Yeast extract	0.20	g	0.199	-
Sodium resazurin (0.1% w/v)	0.50	ml	5.0e-4	0.002
Na₂CO₃	1.50	g	1.496	14.11
Na₂-fumarate	3.20	g	3.19	19.936
Wolin's vitamin solution (10x)	1.00	ml	-	-
Na₂S x 9 H₂O	0.50	g	0.499	2.076
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients (except carbonate, vitamins, fumarate and sulfide) 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 anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving complete the medium by adding vitamins, fumarate and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas and carbonate from a sterile anoxic stock solution prepared under 80% N₂ and 20% CO₂ gas atmosphere. Stock solutions of vitamins and fumarate are sterilized by filtration. Adjust pH of the complete medium to 7.0 - 7.2. After inoculation pressurize culture vials to 0.7 bar overpressure with sterile 80% N₂ and 20% CO₂ gas mixture.
2 Note: A white precipitate forms in this medium after autoclaving, which has however no negative effect on growth.

Compound

Amount

Unit

Conc. [g/L]

Conc. [mM]

Na₂HPO₄ x 2 H₂O

0.53

0.528

3.722

KH₂PO₄

0.41

0.409

3.004

NH₄Cl

0.30

0.299

5.592

CaCl₂ x 2 H₂O

0.11

0.746

MgCl₂ x 6 H₂O

0.10

0.1

0.49

NaCl

0.30

0.299

5.118

Trace element solution SL-10

1.00

Selenite-tungstate solution

1.00

Yeast extract

0.20

0.199

Sodium resazurin
(0.1% w/v)

0.50

5.0e-4

0.002

Na₂CO₃

1.50

1.496

14.11

Na₂-fumarate

3.20

3.19

19.936

Wolin's vitamin solution (10x)

1.00

Na₂S x 9 H₂O

0.50

0.499

2.076

Distilled water

1000.00

1 Dissolve ingredients (except carbonate, vitamins, fumarate and sulfide) 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 anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving complete the medium by adding vitamins, fumarate and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas and carbonate from a sterile anoxic stock solution prepared under 80% N₂ and 20% CO₂ gas atmosphere. Stock solutions of vitamins and fumarate are sterilized by filtration. Adjust pH of the complete medium to 7.0 - 7.2. After inoculation pressurize culture vials to 0.7 bar overpressure with sterile 80% N₂ and 20% CO₂ gas mixture.

2 Note: A white precipitate forms in this medium after autoclaving, which has however no negative effect on growth.

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Main sol. 684

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Main sol. 684

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