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DSMZ

DSMZ 1094 . METHANOSPHAERULA (PEAT) MEDIUM

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

Culture medium recipe

Vol.
×
Medium for Methanosphaerula palustris DSM 19958

This strain uses the standard medium without modifications.

Growth conditions: 7-14 days anaerobic 28 °C

Main sol. 1094 #

ml
Equipment needed: Hungate tubes Gassing station Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 943 ml - -
Solution B 13 ml - -
Solution C 20 ml - -
Solution D 1 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 to 30% of their volume 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. 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. Adjust pH of complete medium to 5.7, if necessary.
2 After inoculation pressurize vials to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
3 Note: The complete medium is not stable and can be stored only for a short time.

Solution A #

943
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl
(0.1% w/v) 		1.5 ml 0.002 0.021
KH2PO4
(0.1% w/v) 		13.6 ml 0.014 0.106
NH4Cl
(0.1% w/v) 		26.8 ml 0.028 0.531
Trace element solution 1.0 ml - -
Distilled water 900.0 ml - -

Solution B #

13
Compound Amount Unit Conc. [g/L] Conc. [mM]
Tris-HCl buffer
(1.0 M solution, pH 8.0) 		7.20 ml - 7.2
Nitrilotriacetate
(0.5 M, neutralized with 10 N NaOH) 		4.80 ml 0.459 2.4
TiCl3
(15% w/v solution in HCl; Riedel-de Haen) 		0.55 ml 6.346 41.15

Solution C #

20
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.

Solution D #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Wolin's vitamin solution (10x) 1 ml - -

Solution E #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
2-Mercaptoethanesulfonate
(Coenzyme M) 		0.08 g 8 56.259
Distilled water 10.00 ml - -

Solution F #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-acetate 0.41 g 41 499.794
Distilled water 10.00 ml - -

Solution G #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 9.6 mg 0.96 3.997
Distilled water 10.0 ml - -

Trace element solution #

1000
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.

Wolin's vitamin solution (10x) #

1000
Solution occurs in 306 other media (first defined in Medium 120).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 20 mg 0.02 0.082
Folic acid 20 mg 0.02 0.045
Pyridoxine hydrochloride 100 mg 0.1 0.486
Thiamine HCl 50 mg 0.05 0.148
Riboflavin 50 mg 0.05 0.133
Nicotinic acid 50 mg 0.05 0.406
Calcium D-(+)-pantothenate 50 mg 0.05 0.21
Vitamin B12 1 mg 1.0e-3 7.4e-4
p-Aminobenzoic acid 50 mg 0.05 0.365
(DL)-alpha-Lipoic acid 50 mg 0.05 0.242
Distilled water 1000 ml - -

Metadata

Last modified: 26.11.24
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
Methanosphaerula palustris DSM 19958

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
2-(N-Morpholino)ethanesulfonic acid 1.936 9.918
Na-acetate 0.407 4.963
NaOH 0.199 4.966
TiCl3 0.082 0.531
2-Mercaptoethanesulfonate 0.079 0.559
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Cultivation metadata from 3 strains BacDive

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