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Main sol. 711a

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Internal Solution-ID: S5648

Equipment needed: Filter Hungate tubes Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.00 g 0.997 18.639
K2HPO4 0.30 g 0.299 1.717
KH2PO4 0.30 g 0.299 2.198
MgCl2 x 6 H2O 0.20 g 0.199 0.981
CaCl2 x 2 H2O 0.10 g 0.1 0.678
KCl 0.10 g 0.1 1.337
NaCl 0.60 g 0.598 10.236
Yeast extract 1.00 g 0.997 -
Fructose 5.00 g 4.985 27.67
Trace element solution SL-10 1.50 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.50 g 1.496 14.11
Wolin's vitamin solution 10.00 ml - -
3,4,5-Trimethoxycinnamic acid
(TMC) 		1.19 g 1.186 4.98
L-Cysteine HCl x H2O 0.30 g 0.299 1.703
Na2S x 9 H2O 0.30 g 0.299 1.245
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, vitamins, TMC and reducing agents) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add vitamins, TMC, 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 atmosphere. The vitamin solution should be sterilized by filtration. The stock solution of TMC (6% w/v) should be neutralized with NaOH prior to sterilization by filtration. Adjust pH of complete medium to 7.0, if necessary.
711a EUBACTERIUM AGGREGANS MEDIUM