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Main sol. 947

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Internal Solution-ID: S1959

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
(NH4)2SO4 0.50 g 0.499 3.773
NH4Cl 0.50 g 0.499 9.32
KH2PO4 2.00 g 1.994 14.653
MgCl2 x 6 H2O 0.04 g 0.04 0.196
CaCl2 x 2 H2O 0.04 g 0.04 0.271
Yeast extract 0.30 g 0.299 -
Glycerol 3.00 g 2.991 32.479
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 1.00 g 0.997 11.868
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.13 g 0.13 0.738
Na2S x 9 H2O 0.13 g 0.13 0.54
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, vitamins and reducing agents), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add solid bicarbonate and equilibrate pH to 6.3 with 80% N2 and 20% CO2 gas mixture. Dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins (sterilized by filtration), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 6.2 - 6.5, if necessary.
947 CALORAMATOR VITERBIENSIS MEDIUM