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119b. METHANOMASSILIICOCCUS MEDIUM

Culture medium recipe

Vol.
×
Equipment needed: Hungate tubes Autoclave
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 0.50 g 0.497 3.652
2 0.40 g 0.398 1.613
3 0.40 g 0.398 6.804
4 0.40 g 0.398 7.433
5 0.05 g 0.05 0.338
6
(0.1% w/v in 0.1 N H2SO4)
2.00 ml 0.002 0.007
7 Trace element solution SL-10 1.00 ml - -
8 Selenite-tungstate solution 1.00 ml - -
9
(OXOID)
1.00 g 0.994 -
10 1.00 g 0.994 12.117
11 2.00 g 1.988 29.233
12 Sludge fluid 50.00 ml - -
13 Fatty acid mixture 20.00 ml - -
14
(0.1% w/v)
0.50 ml 5.0e-4 0.002
15 1.50 ml 1.178 36.762
16 1.00 g 0.994 11.833
17 0.50 g 0.497 2.83
18 0.50 g 0.497 2.069
19 930.00 ml 922.796 51222.9
Instructions:
  1. Dissolve ingredients except methanol, bicarbonate, cysteine and sulfide. Adjust pH of medium to 7.2 and sparge with 100% N2 gas for 30 - 45 min to make it anoxic. Then dispense medium under same gas atmosphere into anoxic Hungate-type tubes to 30% v/v of their volume and autoclave. Add methanol, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use check pH of complete medium and adjust to 7.6, if necessary. After inoculation, add sterile 80% H2 and 20% CO2 gas mixture to 0.5 bar overpressure. After growth becomes visible overpressure of 80% H2 and 20% CO2 gas mixture can be increased to 1 bar.
Solution occurs in 161 other media (first defined in medium 385 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 0.5 g 0.5 12.501
2 3.0 mg 0.003 0.011
3 4.0 mg 0.004 0.012
4 1000.0 ml 998.207 55408.8
Solution occurs in 8 other media (first defined in medium 119 )
Equipment needed: Autoclave Anaerobic workbench Centrifuge
Instructions:
  1. Add 0.4% yeast extract to sludge from an anaerobic digester, and after gassing with nitrogen gas for a few minutes incubate it at 37°C for 24 hours Then centrifuge the sludge at 13000 g and autoclave the resulting, clear supernatant in screw-capped vessels under nitrogen gas. The sludge fluid can be stored at room temperature in the dark.
Solution occurs in 10 other media (first defined in medium 119 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 23 ml - -
2 27 ml - -
3 27 ml 22.989 225.092
4 27 ml - -
5 1000 ml 904.173 50189.2
Instructions:
  1. Adjust pH to 7.0 - 7.3 with 2 N NaOH.
Solution occurs in 239 other media (first defined in medium 320 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
2
(25%; 7.7 M)
10.0 ml 3.17 86.943
3 1.5 g 1.5 7.545
4 70.0 mg 0.07 0.514
5 100.0 mg 0.1 0.505
6 6.0 mg 0.006 0.097
7 190.0 mg 0.19 0.799
8 2.0 mg 0.002 0.012
9 24.0 mg 0.024 0.101
10 36.0 mg 0.036 0.149
11 990.0 ml 988.225 54854.8
Instructions:
  1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.0 ml.

Metadata

Media status: Official DSMZ medium
Source: DSMZ
Taxonomic range:  Archaea
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.6
Equipment needed: Anaerobic workbench, Autoclave, Centrifuge, Hungate tubes

Associated strains

All strains for this medium: 1
Strains with modifications: 1

Gas composition

  • 20 % CO2
  • 80 % H2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Distilled water 942.746 52330.3
Sodium formate 1.988 29.233
Methanol 1.178 36.762
Sodium acetate 0.994 12.117
NaHCO3 0.994 11.833
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Cultivation metadata from 1 strains BacDive

halophily not 1 strain
pH 6 - 8 1 strain
phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain

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