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DSMZ

DSMZ 825a . METHANOBACTERIUM II MEDIUM (N2/CO2)

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

Culture medium recipe

Vol.
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Medium for Methanosarcina mazei DSM 10132

This strain uses the standard medium without modifications.

Growth conditions: anaerobic 37 °C

Main sol. 825a #

ml
Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.0 g 0.979 18.311
K2HPO4 0.3 g 0.294 1.687
KH2PO4 0.3 g 0.294 2.159
MgCl2 x 6 H2O 0.2 g 0.196 0.964
CaCl2 x 2 H2O 0.1 g 0.098 0.666
KCl 0.1 g 0.098 1.314
NaCl 0.6 g 0.588 10.056
Modified Wolin's mineral solution 10.0 ml - -
Na-acetate 0.5 g 0.49 5.97
Yeast extract 1.0 g 0.979 -
Sodium resazurin
(0.1% w/v) 		0.5 ml 4.9e-4 0.002
NaHCO3 4.0 g 3.918 46.636
Methanol
(15% v/v) 		10.0 ml 7.757 242.092
Wolin's vitamin solution (10x) 1.0 ml - -
L-Cysteine HCl x H2O 0.5 g 0.49 2.788
Na2S x 9 H2O 0.5 g 0.49 2.039
Distilled water 1000.0 ml - -
1 Dissolve ingredients except bicarbonate, methanol, vitamins, cysteine and sulfide. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate, adjust pH to 7.0, then dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add methanol (15% v/v solution), vitamins (sterilized by filtration), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to use check pH of complete medium and adjust to 6.8 - 7.0, if necessary.

Modified Wolin's mineral solution #

1000
Solution occurs in 167 other media (first defined in Medium 141).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid 1.50 g 1.5 7.848
MgSO4 x 7 H2O 3.00 g 3 12.172
MnSO4 x H2O 0.50 g 0.5 2.958
NaCl 1.00 g 1 17.112
FeSO4 x 7 H2O 0.10 g 0.1 0.36
CoSO4 x 7 H2O 0.18 g 0.18 1.161
CaCl2 x 2 H2O 0.10 g 0.1 0.68
ZnSO4 x 7 H2O 0.18 g 0.18 0.626
CuSO4 x 5 H2O 0.01 g 0.01 0.04
AlK(SO4)2 x 12 H2O 0.02 g 0.02 0.042
H3BO3 0.01 g 0.01 0.162
Na2MoO4 x 2 H2O 0.01 g 0.01 0.041
NiCl2 x 6 H2O 0.03 g 0.03 0.231
Na2SeO3 x 5 H2O 0.30 mg 3.0e-4 0.001
Na2WO4 x 2 H2O 0.40 mg 4.0e-4 0.001
Distilled water 1000.00 ml - -
1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.

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: 02.12.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 6.8 - 7.0
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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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: 3
Strains with modifications: 2
Methanosarcina mazei DSM 10132

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Methanol 7.757 242.092
NaHCO3 3.918 46.636
NH4Cl 0.979 18.311
Yeast extract 0.979 -
Na-acetate 0.49 5.97
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Cultivation metadata from 3 strains BacDive

phyla 1 3 strains
oxygen anaerobe 3 strains
temperature mesophilic 3 strains