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Main sol. 298b

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Internal Solution-ID: S566

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.20 g 0.2 1.467
NH4Cl 0.25 g 0.25 4.664
NaCl 1.00 g 0.998 17.077
MgCl2 x 6 H2O 0.40 g 0.399 1.964
KCl 0.50 g 0.499 6.694
CaCl2 x 2 H2O 0.15 g 0.15 1.018
Trace element solution SL-10 1.00 ml - -
Na-acetate 0.20 g 0.2 2.433
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.50 g 1.497 14.124
Na2-glyoxalate 0.60 g 0.599 6.236
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.299 1.705
Na2S x 9 H2O 0.30 g 0.299 1.247
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonate, glyoxalate, 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 in anoxic Hungate-type tubes or serum vials under the same gas atmosphere and autoclave. Add glyoxalate (sterilized by filtration), vitamins (sterilized by filtration), cysteine 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 mixture. Prior to use adjust pH of complete medium to 7.2 - 7.5.
2 Note: When growth has started, feed culture once or twice with the same amount of glyoxalate.
298b SYNTROPHOBOTULUS MEDIUM