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Main sol. 1616

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Internal Solution-ID: S3363

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 20.00 g 19.92 340.868
NH4Cl 0.54 g 0.538 10.055
KH2PO4 0.10 g 0.1 0.732
MgCl2 x 7 H2O 4.00 g 3.984 41.845
CaCl2 x 2 H2O 1.00 g 0.996 6.775
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Yeast extract 1.00 g 0.996 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.50 g 1.494 14.096
Na-pyruvate 2.75 g 2.739 24.89
Wolin's vitamin solution (10x) 2.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.299 1.701
Na2S x 9 H2O 0.30 g 0.299 1.244
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonate, pyruvate, vitamins, cysteine and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Distribute medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add pyruvate, vitamins, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. The pyruvate and vitamin solutions should be sterilized by filtration. Check pH of complete medium and adjust to 7.2, if necessary.
1616 SEDIMENTIBACTER ACIDAMINIVORANS MEDIUM