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Main sol. 1004

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Internal Solution-ID: S2050

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.14 g 0.14 1.026
MgCl2 x 6 H2O 0.20 g 0.199 0.981
CaCl2 x 2 H2O 0.15 g 0.15 1.017
NH4Cl 0.54 g 0.538 10.065
Trace element solution SL-11 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 2.50 g 2.493 29.67
Yeast extract 2.30 g 2.293 -
D-Glucose 2.20 g 2.193 12.175
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.25 g 0.249 1.419
Na2S x 9 H2O 0.25 g 0.249 1.038
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, yeast extract, vitamins, glucose 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 adjust pH to 7.0. Dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add yeast extract, vitamins (sterilized by filtration), glucose, sulfide and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 7.0, if necessary.
1004 ANAEROLINEA MEDIUM