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Main sol. 500

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Internal Solution-ID: S1027

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 0.35 g 0.348 1.999
KH2PO4 0.23 g 0.229 1.682
NH4Cl 0.50 g 0.498 9.301
NaCl 2.25 g 2.239 38.31
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) 		2.00 ml 0.002 0.007
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Yeast extract 1.00 g 0.995 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 3.00 g 2.985 35.534
MgSO4 x 7 H2O 0.50 g 0.498 2.019
CaCl2 x 2 H2O 0.25 g 0.249 1.692
D-Glucose 5.00 g 4.975 27.615
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.299 1.7
Na2S x 9 H2O 0.30 g 0.299 1.243
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, magnesium sulfate, calcium chloride, glucose, vitamins, cysteine and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate, adjust pH to 7.0, dispense under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add magnesium sulfate, calcium chloride, glucose, cysteine and sulfide from sterile anoxic stock solutions autoclaved under 100% N2 gas. Vitamins are prepared under 100% N2 gas atmosphere and sterilized by filtration. Adjust pH of the complete medium to 7.0 - 7.2, if necessary.
500 CLOSTRIDIUM (GV) MEDIUM