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Main sol. 380

Internal Solution-ID: S723

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
KH₂PO₄	0.68	g	0.68	4.997
NaNO₃	0.12	g	0.12	1.412
L(+)-Tartaric acid	0.37	g	0.37	2.465
Succinic acid	0.37	g	0.37	3.133
Na-acetate	0.05	g	0.05	0.61
Yeast extract	0.10	g	0.1	-
Modified Wolin's mineral solution	5.00	ml	-	-
Fe(III) quinate solution 0.01 M	2.00	ml	-	-
Agar , for semi-solid medium (BD Bacto) optional	1.30	g	1.3	3.865
Sodium resazurin (0.1% w/v)	0.50	ml	5.0e-4	-
Na-thioglycolate	0.05	g	0.05	0.438
Seven vitamins solution	1.00	ml	-	-
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients (except thioglycolate and vitamins) in the order given and adjust pH to 6.75 with NaOH.
2 Preparation of liquid medium: Sparge medium with 100% N₂ gas for 30 -45 min and dispense under the same gas atmosphere into anoxic Hungate-type tubes to 50% of their volume. Seal vials with screw caps and gas tight butyl rubber closures. Autoclave at 121°C for 15 min. Before inoculation add thioglycolate and vitamins from stock solutions prepared under 100% N₂ gas and filter-sterilized. Then add sterile air (with hypodermic syringe through the rubber closure) to a concentration of ca. 1% (v/v) O₂ in the vial (e.g., add 1 ml air to a Hungate-type tube of 16 ml total volume).
3 Preparation of semi solid medium: Supplement medium with agar, bring medium to the boil and cool under 100% N₂ gas atmosphere. Dispense under same gas atmosphere aliquots of 10 ml semi-solid medium into Hungate-type tubes. Prior to inoculation add thioglycolate from a 0.5% (w/v) stock solution, freshly prepared under 100% N₂ gas and filter-sterilized. Then add sterile air (with hypodermic syringe through the rubber closure) to a concentration of ca. 1% (v/v) in the vial.
4 Note: Prior to inoculation media should be slightly pink in color. Strongly reduced conditions will not support growth of microaerophilic Magnetospirillum species. Use as inoculum 10% (v/v). Incubate tubes with medium without agitation in an inclined position. During growth O₂ will be consumed and the pH will increase. For cultivation of magnetic cells we recommend preparation of liquid medium, while semi-solid medium is more suitable for demonstration of microaerophilic band formation and storage.

Compound

Amount

Unit

Conc. [g/L]

Conc. [mM]

KH₂PO₄

0.68

4.997

NaNO₃

0.12

1.412

L(+)-Tartaric acid

0.37

2.465

Succinic acid

0.37

3.133

Na-acetate

0.05

0.61

Yeast extract

0.10

0.1

Modified Wolin's mineral solution

5.00

Fe(III) quinate solution 0.01 M

2.00

Agar , for semi-solid medium
(BD Bacto)
optional

1.30

1.3

3.865

Sodium resazurin
(0.1% w/v)

0.50

5.0e-4

Na-thioglycolate

0.05

0.438

Seven vitamins solution

1.00

Distilled water

1000.00

1 Dissolve ingredients (except thioglycolate and vitamins) in the order given and adjust pH to 6.75 with NaOH.

2 Preparation of liquid medium: Sparge medium with 100% N₂ gas for 30 -45 min and dispense under the same gas atmosphere into anoxic Hungate-type tubes to 50% of their volume. Seal vials with screw caps and gas tight butyl rubber closures. Autoclave at 121°C for 15 min. Before inoculation add thioglycolate and vitamins from stock solutions prepared under 100% N₂ gas and filter-sterilized. Then add sterile air (with hypodermic syringe through the rubber closure) to a concentration of ca. 1% (v/v) O₂ in the vial (e.g., add 1 ml air to a Hungate-type tube of 16 ml total volume).

3 Preparation of semi solid medium: Supplement medium with agar, bring medium to the boil and cool under 100% N₂ gas atmosphere. Dispense under same gas atmosphere aliquots of 10 ml semi-solid medium into Hungate-type tubes. Prior to inoculation add thioglycolate from a 0.5% (w/v) stock solution, freshly prepared under 100% N₂ gas and filter-sterilized. Then add sterile air (with hypodermic syringe through the rubber closure) to a concentration of ca. 1% (v/v) in the vial.

4 Note: Prior to inoculation media should be slightly pink in color. Strongly reduced conditions will not support growth of microaerophilic Magnetospirillum species. Use as inoculum 10% (v/v). Incubate tubes with medium without agitation in an inclined position. During growth O₂ will be consumed and the pH will increase. For cultivation of magnetic cells we recommend preparation of liquid medium, while semi-solid medium is more suitable for demonstration of microaerophilic band formation and storage.

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Main sol. 380

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Main sol. 380

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