Back to all solutions

Main sol. 141a

Internal Solution-ID: S243

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.34 g 0.333 4.463
MgCl2 x 6 H2O 4.00 g 3.914 19.252
MgSO4 x 7 H2O 3.45 g 3.376 13.696
NH4Cl 0.25 g 0.245 4.573
CaCl2 x 2 H2O 0.14 g 0.137 0.932
K2HPO4 0.14 g 0.137 0.786
NaCl 18.00 g 17.613 301.378
Modified Wolin's mineral solution 10.00 ml - -
Fe(NH4)2(SO4)2 x 6 H2O
(0.1% w/v)
2.00 ml 0.002 -
Na-acetate 1.00 g 0.978 11.928
Sodium resazurin
(0.1% w/v)
0.50 ml 4.9e-4 0.002
Wolin's vitamin solution 10.00 ml - -
L-Cysteine HCl x H2O 0.50 g 0.489 2.786
Na2S x 9 H2O 0.50 g 0.489 2.037
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except vitamins, cysteine and sulfide), sparge medium with 80% H2 and 20% CO2 gas mixture to reach a pH below 6 (at least 30 min), then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. After sterilization add cysteine and sulfide from sterile anoxic stock solutions autoclaved under 100% N2 gas atmosphere. Vitamins are prepared under 100% N2 gas atmosphere and sterilized by filtration. Adjust pH of final medium to 5.5 - 5.7.
2 For incubation use sterile 80% H2 and 20% CO2 gas mixture at two atmospheres of pressure.
141a METHANOCOCCUS SP. MEDIUM