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Main sol. 671

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Internal Solution-ID: S1420

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.00 g 0.989 18.492
NaCl 0.10 g 0.099 1.693
MgCl2 x 6 H2O 0.10 g 0.099 0.487
CaCl2 x 2 H2O 0.05 g 0.049 0.336
K2HPO4 x 3 H2O 0.40 g 0.396 2.272
Modified Wolin's mineral solution 10.00 ml - -
Yeast extract 0.75 g 0.742 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
Na2CO3 1.00 g 0.989 9.332
Cellobiose 4.00 g 3.956 11.559
Cellulose , Avicel SIGMA
optional 		2.00 g 1.978 -
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.25 g 0.247 1.03
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, cellobiose, vitamins and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add cellobiose, vitamins 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. Stock solutions of cellobiose and vitamins should be sterilized by filtration. Adjust pH of complete medium to 7.0, if necessary.
2 Note: Some strains can be adapted to cellulose as substrate using 2.00 g/l Avicel microcrystalline cellulose (SIGMA).
671 THERMOANAEROBACTER (BA) MEDIUM