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Main sol. 558

Internal Solution-ID: S1219

Equipment needed: Hungate tubes Filter Gassing station Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 1.50 g 1.484 10.903
Na2HPO4 x 12 H2O 4.20 g 4.154 29.264
NH4Cl 0.50 g 0.495 9.246
MgCl2 x 6 H2O 0.36 g 0.356 1.751
Na2SO4 1.00 g 0.989 6.964
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 4.9e-4 0.002
D-Glucose 10.00 g 9.891 54.902
Yeast extract 2.00 g 1.978 -
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.50 g 0.495 2.816
Na2S x 9 H2O 0.50 g 0.495 2.059
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except glucose, yeast extract, vitamin solution and reducing agents), adjust pH to 3.5 using HCl and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials. After autoclaving add glucose, yeast extract, vitamin solution, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The vitamin solution should be sterilized by filtration. Adjust pH of complete medium to 3.5, if necessary.
558 CLOSTRIDIUM (RB-1) MEDIUM