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Main sol. 201

Internal Solution-ID: S415

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Solution A	930	ml	-	-
Solution B	1	ml	-	-
Solution C	30	ml	-	-
Solution D	10	ml	-	-
Solution E	1	ml	-	-
Solution F	1	ml	-	-
Solution G	20	ml	-	-
Solution H	10	ml	-	-
1 Solution A is sparged with 80% N₂ and 20% CO₂ gas mixture for 30 - 45 min to reach a pH of around 6, then distributed under same gas atmosphere into anoxic Hungate-type tubes and autoclaved.
2 Solutions B, D, E, G, and H are autoclaved separately under 100% N₂ gas. Solution C is autoclaved under 80% N₂ and 20% CO₂ gas atmosphere. Solution F is prepared under 100% N₂ gas and sterilized by filtration. To complete the medium appropriate amounts of solutions B to H are added to the sterile solution A in the sequence as indicated. Adjust pH of the complete medium to 7.6, if necessary.
3 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution freshly prepared under N₂ and filter-sterilized) just before inoculation may stimulate growth at the beginning. For transfers use 5 - 10% inoculum.
4 Cultures of Desulfonema spp. show usually improved growth in media containing an artificial light sediment of aluminum phosphate. Alternatively, a viscous medium prepared with 1 - 2 g washed agar per liter can be used (Widdel et al., Arch. Microbiol. 134:286-294 (1983)): Agar (Difco) is washed three times for one hour with distilled water at room temperature. Finally, the agar is suspended (c. 20.0 g/l) in distilled water and autoclaved. 50 ml of the hot agar is added to the hot sterile medium part A (when preparing part A calculated for 1 liter, subtract the volume of the agar solution). After cooling to room temperature complete the medium by adding part B to I. For mass cultures the aluminum phosphate as artificial sediment can be used.

Compound

Amount

Unit

Conc. [g/L]

Conc. [mM]

Solution A

930

Solution B

Solution C

Solution D

Solution E

Solution F

Solution G

Solution H

1 Solution A is sparged with 80% N₂ and 20% CO₂ gas mixture for 30 - 45 min to reach a pH of around 6, then distributed under same gas atmosphere into anoxic Hungate-type tubes and autoclaved.

2 Solutions B, D, E, G, and H are autoclaved separately under 100% N₂ gas. Solution C is autoclaved under 80% N₂ and 20% CO₂ gas atmosphere. Solution F is prepared under 100% N₂ gas and sterilized by filtration. To complete the medium appropriate amounts of solutions B to H are added to the sterile solution A in the sequence as indicated. Adjust pH of the complete medium to 7.6, if necessary.

3 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution freshly prepared under N₂ and filter-sterilized) just before inoculation may stimulate growth at the beginning. For transfers use 5 - 10% inoculum.

4 Cultures of Desulfonema spp. show usually improved growth in media containing an artificial light sediment of aluminum phosphate. Alternatively, a viscous medium prepared with 1 - 2 g washed agar per liter can be used (Widdel et al., Arch. Microbiol. 134:286-294 (1983)): Agar (Difco) is washed three times for one hour with distilled water at room temperature. Finally, the agar is suspended (c. 20.0 g/l) in distilled water and autoclaved. 50 ml of the hot agar is added to the hot sterile medium part A (when preparing part A calculated for 1 liter, subtract the volume of the agar solution). After cooling to room temperature complete the medium by adding part B to I. For mass cultures the aluminum phosphate as artificial sediment can be used.

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Main sol. 201

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Main sol. 201

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