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Main sol. 1051

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Internal Solution-ID: S2127

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.14 g 0.14 1.029
MgCl2 x 6 H2O 0.20 g 0.2 0.984
CaCl2 x 2 H2O 0.15 g 0.15 1.02
NH4Cl 0.54 g 0.54 10.095
Trace element solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 -
Na2CO3 1.50 g 1.5 14.153
Yeast extract 2.30 g 2.3 -
Wolin's vitamin solution 2.00 ml - -
D-Glucose 2.20 g 2.2 12.211
L-Cysteine HCl x H2O 0.30 g 0.3 1.708
Na2S x 9 H2O 0.30 g 0.3 1.249
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, yeast extract, vitamins, glucose, cysteine and sulfide), then 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. Add yeast extract, vitamins (sterilized by filtration), glucose, 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. Adjust pH of the complete medium to 7.0-7.2, if necessary.
1051 TEPIDANAEROBACTER MEDIUM