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Main sol. 557

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Internal Solution-ID: S1218

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
CaSO4 x 2 H2O 0.05 g 0.05 0.29
MgSO4 x 7 H2O 0.50 g 0.499 2.023
FeSO4 x 7 H2O 0.01 g 0.01 0.036
MnCl2 x 4 H2O 0.06 g 0.06 0.302
NaCl 1.00 g 0.997 17.06
Trace element solution SL-11 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Yeast extract 5.00 g 4.985 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
KH2PO4 2.64 g 2.632 19.342
K2HPO4 5.33 g 5.314 30.51
NaHCO3 1.00 g 0.997 11.868
Creatinine 5.50 g 5.484 48.476
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.50 g 0.499 2.838
Na2S x 9 H2O 0.50 g 0.499 2.076
Distilled water 1000.00 ml - -
1 Dissolve ingredients except hydrogenphosphates, bicarbonate, creatinine, vitamins, cysteine and sulfide, adjust pH to 7.0 and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Complete medium by adding hydrogenphosphates, creatinine, vitamins, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Stock solutions of creatinine (5% w/v) and vitamins should be sterilized by filtration. Adjust pH of complete medium to 7.0, if necessary.
557 CLOSTRIDIUM SP. MEDIUM (CREATININE)