DSMZ 1094 . METHANOSPHAERULA (PEAT) MEDIUM

Official DSMZ medium: This medium was manually curated by experts from the DSMZ.

Culture medium recipe

Vol.
×
Equipment needed: Hungate tubes Gassing station Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 933 ml - -
Solution B 13 ml - -
Solution C 20 ml - -
Solution D 10 ml - -
Solution E 10 ml - -
Solution F 10 ml - -
Solution G 10 ml - -
1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make anoxic. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solutions B, C, D and G are prepared under 100% N2 gas atmosphere and sterilized by filtration. Solutions E and F are prepared under 100% N2 gas atmosphere and autoclaved.
2 To complete the medium add appropriate amounts of solutions B to G to the sterile solution A in the sequence indicated and let medium equilibrate overnight. After inoculation pressurize vials to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture. Adjust pH of complete medium to 5.7, if necessary.
3 Note: The complete medium is not stable and can be stored only for a short time.
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl
(0.1% w/v)
1.5 ml 0.002 0.02
KH2PO4
(0.1% w/v)
13.6 ml 0.014 0.1
NH4Cl
(0.1% w/v)
26.8 ml 0.027 0.501
Trace element solution 1.0 ml - -
Distilled water 890.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Tris-HCl buffer , pH 8.0)
(1.0 M solution, pH 8.0, 1 M)
7.20 ml - 7.2
Na3-NTA x H2O
(0.5 M solution, 0.5 M)
4.80 ml 0.511 2.4
TiCl3
(15% w/v solution in HCl; Riedel-de Haen)
0.55 ml 0.083 0.535
Compound Amount Unit Conc. [g/L] Conc. [mM]
2-(N-Morpholino)ethanesulfonic acid
(MES)
1.95 g 97.5 499.385
NaOH 0.20 g 10 250.019
Distilled water 20.00 ml - -
1 Adjust to pH 7.5 with NaOH.
Compound Amount Unit Conc. [g/L] Conc. [mM]
Wolin's vitamin solution 10 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
2-Mercaptoethanesulfonic acid
(Coenzyme M)
0.08 g 8 56.259
Distilled water 10.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-acetate 0.41 g 41 499.794
Distilled water 10.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 9.6 mg 0.96 3.997
Distilled water 10.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2-EDTA 37.23 g 37.23 110.736
FeCl2 x 4 H2O 1.34 g 1.34 6.74
AlK(SO4)2 x 12 H2O 3.45 g 3.45 7.273
CaCl2 x 2 H2O 2.34 g 2.34 15.917
MgSO4 x 7 H2O 1.56 g 1.56 6.329
CoCl2 x 6 H2O 24.00 mg 0.024 0.185
ZnCl2 75.00 mg 0.075 0.55
H3BO3 19.00 mg 0.019 0.307
NiCl2 x 6 H2O 24.00 mg 0.024 0.185
Na2Mo4 x 2 H2O 24.00 mg 0.024 -
MnSO4 x 4 H2O 26.00 mg 0.026 0.117
CuSO4 x 5 H2O 9.00 mg 0.009 0.036
Distilled water 1000.00 ml - -
1 Dissolve EDTA in distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add other compounds.
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: 12.04.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Defined medium
Final pH: 5.7
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 40 % N2
  • 40 % H2
  • 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: 3
Strains with modifications: 2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
2-(N-Morpholino)ethanesulfonic acid 1.938 9.928
Na-acetate 0.408 4.968
NaOH 0.199 4.971
2-Mercaptoethanesulfonic acid 0.08 0.559
Na2-EDTA 0.037 0.11
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Cultivation metadata from 3 strains BacDive

phyla 1 3 strains
oxygen anaerobe 2 strains
temperature mesophilic 3 strains
temperature psychrophilic 1 strain