Main sol. 769
Internal Solution-ID: S1567
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Sea Salt
(SIGMA) |
12.0 | g | 11.988 | - | |
NH4NO3 | 0.1 | g | 0.1 | - | |
Yeast extract
(BD Bacto) |
1.0 | g | 0.999 | - | |
Trypticase peptone
(BD BBL) |
0.4 | g | 0.4 | - | |
Sodium resazurin
(0.1% w/v) |
0.5 | ml | 5.0e-4 | 0.002 | |
NaHCO3 | 1.0 | g | 0.999 | 11.892 | |
Na-phosphate buffer
(0.2 M, pH 6.5) |
20.0 | ml | - | 4 | |
Lactose | 2.0 | g | 1.998 | 5.837 | |
Wolin's vitamin solution (10x) | 1.0 | ml | - | - | |
L-Cysteine HCl x H2O | 0.3 | g | 0.3 | 1.706 | |
Na2S x 9 H2O | 0.3 | g | 0.3 | 1.248 | |
Distilled water | 980.0 | ml | - | - | |
1 Dissolve ingredients (except bicarbonate, phosphate-buffer, lactose, vitamins, cysteine and sulfide), then 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 and autoclave. After sterilization add phosphate-buffer, lactose, cysteine and sulfide from sterile anoxic stock solutions autoclaved under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Vitamins are prepared under N2 gas atmosphere and sterilized by filtration. Adjust pH of complete medium to 6.5, if necessary. | |||||
2 Note: Sea salts can be replaced by 300.0 ml/l of filtered aged sea water. |
769 | CLOSTRIDIUM VINCENTII MEDIUM |