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Main sol. 1527a

Internal Solution-ID: S3166

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
NaCl	90.0	g	89.021	1523.28
MgCl₂ x 6 H₂O	6.0	g	5.935	29.192
KCl	1.5	g	1.484	19.902
Na₂SO₄	1.0	g	0.989	6.964
NH₄Cl	1.0	g	0.989	18.492
CaCl₂ x 2 H₂O	0.4	g	0.396	2.691
K₂HPO₄	0.4	g	0.396	2.272
Modified Wolin's mineral solution	10.0	ml	-	-
Yeast extract (BD Bacto)	1.0	g	0.989	-
Sodium resazurin (0.1% w/v)	0.5	ml	4.9e-4	0.002
Na₂CO₃	1.5	g	1.484	13.999
Trypticase peptone (BD BBL)	1.0	g	0.989	-
D-Glucose	1.0	g	0.989	5.49
Wolin's vitamin solution (10x)	1.0	ml	-	-
L-Cysteine HCl x H₂O	0.5	g	0.495	2.816
Na₂S x 9 H₂O	0.5	g	0.495	2.059
Distilled water	1000.0	ml	-	-
1 Dissolve ingredients except carbonate, peptone, glucose, vitamins, cysteine and sulfide, then sparge medium with 80% N₂ and 20% CO₂ gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add peptone, glucose, vitamins, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas (vitamins are sterilized by filtration) and carbonate from a sterile anoxic stock solution prepared under 80% N₂ and 20% CO₂ gas mixture. Prior to use adjust pH of complete medium to 7.3 - 7.5, if necessary.
2 Note: Use at least 5-10% (v/v) inoculum.

Compound

Amount

Unit

Conc. [g/L]

Conc. [mM]

NaCl

90.0

89.021

1523.28

MgCl₂ x 6 H₂O

6.0

5.935

29.192

KCl

1.5

1.484

19.902

Na₂SO₄

1.0

0.989

6.964

NH₄Cl

1.0

0.989

18.492

CaCl₂ x 2 H₂O

0.4

0.396

2.691

K₂HPO₄

0.4

0.396

2.272

Modified Wolin's mineral solution

10.0

Yeast extract
(BD Bacto)

1.0

0.989

Sodium resazurin
(0.1% w/v)

0.5

4.9e-4

0.002

Na₂CO₃

1.5

1.484

13.999

Trypticase peptone
(BD BBL)

1.0

0.989

D-Glucose

1.0

0.989

5.49

Wolin's vitamin solution (10x)

1.0

L-Cysteine HCl x H₂O

0.5

0.495

2.816

Na₂S x 9 H₂O

0.5

0.495

2.059

Distilled water

1000.0

1 Dissolve ingredients except carbonate, peptone, glucose, vitamins, cysteine and sulfide, then sparge medium with 80% N₂ and 20% CO₂ gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add peptone, glucose, vitamins, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas (vitamins are sterilized by filtration) and carbonate from a sterile anoxic stock solution prepared under 80% N₂ and 20% CO₂ gas mixture. Prior to use adjust pH of complete medium to 7.3 - 7.5, if necessary.

2 Note: Use at least 5-10% (v/v) inoculum.

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Main sol. 1527a

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Main sol. 1527a

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