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Main sol. 212a

Internal Solution-ID: S452

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Solution A	917	ml	-	-
Solution B	30	ml	-	-
Solution C	4	ml	-	-
Solution D	4	ml	-	-
Solution E	30	ml	-	-
Solution F	1	ml	-	-
Solution G	2	ml	-	-
Solution H	10	ml	-	-
Solution I	10	ml	-	-
1 Add and dissolve ingredients of solution A, adjust pH to 7.0, and 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 to 30% of their volume and autoclave. Solution B is prepared under 80% N₂ and 20% CO₂ gas atmosphere and autoclaved. Solutions C, D, and H are autoclaved under a 100% N₂ gas atmosphere. Solutions E, F, G, and I are prepared under 100% N₂ gas and sterilized by filtration. To complete the medium appropriate amounts of solutions B to I are added to the sterile solution A in the sequence as indicated.
2 Note: Some cultures are shipped in semi-solid medium which stimulates growth at the beginning. For agar stabs 3.00 g/l agar are added to the complete medium from a sterile anoxic stock solution (2% w/v). Upon receipt add anoxically 1 - 2 ml of the recommended freshly prepared liquid medium to the agar tube and incubate for 3 - 5 days . After incubation transfer 0.5 ml of the resulting cell suspension in the liquid phase to tubes with liquid medium.

Compound

Amount

Unit

Conc. [g/L]

Conc. [mM]

Solution A

917

Solution B

Solution C

Solution D

Solution E

Solution F

Solution G

Solution H

Solution I

1 Add and dissolve ingredients of solution A, adjust pH to 7.0, and 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 to 30% of their volume and autoclave. Solution B is prepared under 80% N₂ and 20% CO₂ gas atmosphere and autoclaved. Solutions C, D, and H are autoclaved under a 100% N₂ gas atmosphere. Solutions E, F, G, and I are prepared under 100% N₂ gas and sterilized by filtration. To complete the medium appropriate amounts of solutions B to I are added to the sterile solution A in the sequence as indicated.

2 Note: Some cultures are shipped in semi-solid medium which stimulates growth at the beginning. For agar stabs 3.00 g/l agar are added to the complete medium from a sterile anoxic stock solution (2% w/v). Upon receipt add anoxically 1 - 2 ml of the recommended freshly prepared liquid medium to the agar tube and incubate for 3 - 5 days . After incubation transfer 0.5 ml of the resulting cell suspension in the liquid phase to tubes with liquid medium.

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Main sol. 212a

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Main sol. 212a

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