DSMZ 334 . METHANOTHRIX MEDIUM

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Culture medium recipe

Vol.
×
Equipment needed: Gassing station Serum bottle Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 962 ml - -
Solution B 25 ml - -
Solution C 10 ml - -
Solution D 10 ml - -
Solution E 10 ml - -
Solution F 10 ml - -
1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, dispense under same gas atmosphere into anoxic serum vials (e.g., 20 ml medium in 50 ml bottles) and autoclave. Prior to inoculation complete the medium by adding appropriate amounts of sterile solutions C, D, E and F prepared under 100% N2 gas and solution B prepared under 80% N2 and 20% CO2 gas atmosphere. Vitamins are sterilized by filtration. Adjust the pH of the complete medium to 7.2.
2 Note: Use 20% (v/v) inoculum.
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.22 g 0.229 1.681
Na2HPO4 x 2 H2O 0.86 g 0.894 6.297
Na-acetate 6.80 g 7.069 86.167
Na2-EDTA
(0.1% w/v)
1.00 ml 1.0e-3 0.003
Trace elements solution 10.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
Distilled water 950.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1.25 g 50 471.752
Distilled water 25.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 0.30 g 30 513.347
CaCl2 x 2 H2O 0.11 g 11 74.822
MgCl2 x 6 H2O 0.10 g 10 49.188
Distilled water 10.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.3 g 30 560.852
Distilled water 10.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Wolin's vitamin solution 10 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 0.5 g 50 208.175
Distilled water 10.0 ml - -
Solution occurs in 1 other media (first defined in Medium 334).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid
(NTA)
12.80 g 12.8 66.967
FeCl3 x 6 H2O 1.35 g 1.35 4.995
MnCl2 x 4 H2O 0.10 g 0.1 0.505
CoCl2 x 6 H2O 0.03 g 0.03 0.231
CaCl2 x 2 H2O 0.10 g 0.1 0.68
ZnCl2 0.10 g 0.1 0.734
CuCl2 0.03 g 0.03 0.223
H3BO3 0.01 g 0.01 0.162
Na2MoO4 x 2 H2O 0.03 g 0.03 0.124
NiCl2 x 6 H2O 0.12 g 0.12 0.926
NaCl 1.00 g 1 17.112
Na2SeO3 x 5 H2O 0.03 g 0.03 0.114
Distilled water 1000.00 ml - -
1 First dissolve NTA in 200 ml of distilled water and adjust pH to 6.5 with KOH, then dissolve mineral salts. Finally adjust pH to 6.5 with KOH and make up to 1000.00 ml.
Solution occurs in 357 other media (first defined in Medium 141).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 2.0 mg 0.002 0.008
Folic acid 2.0 mg 0.002 0.005
Pyridoxine hydrochloride 10.0 mg 0.01 0.049
Thiamine HCl 5.0 mg 0.005 0.015
Riboflavin 5.0 mg 0.005 0.013
Nicotinic acid 5.0 mg 0.005 0.041
Calcium D-(+)-pantothenate 5.0 mg 0.005 -
Vitamin B12 0.1 mg 1.0e-4 7.4e-5
p-Aminobenzoic acid 5.0 mg 0.005 0.036
(DL)-alpha-Lipoic acid 5.0 mg 0.005 0.024
Distilled water 1000.0 ml - -

Metadata

Last modified: 31.03.22
Source: DSMZ
Medium type: Defined medium
Final pH: 7.2
Equipment needed: Autoclave, Filter, Gassing station, Serum bottle
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Gas composition

  • 80 % N2
  • 20 % CO2

Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .

Associated strains

All strains for this medium: No strains found

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na-acetate 6.8 82.893
Na2CO3 1.25 11.794
Na2HPO4 x 2 H2O 0.86 6.058
Na2S x 9 H2O 0.5 2.082
NH4Cl 0.3 5.609
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Cultivation metadata from 0 strains BacDive

pH 6 - 8 1 strain
pH 8 - 10 1 strain
phyla 1 3 strains
oxygen anaerobe 3 strains
temperature mesophilic 3 strains