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DSMZ

DSMZ 814 . METHANOBACTERIUM SUBTERRANEUM MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 814 #

ml
Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.40 g 0.4 7.478
MgCl2 x 6 H2O 0.03 g 0.03 0.148
NaCl 0.45 g 0.45 7.7
K2HPO4 0.50 g 0.5 2.871
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) 		3.00 ml 0.003 0.011
Modified Wolin's mineral solution 10.00 ml - -
Tryptone
(BD Bacto) 		1.00 g 1 14.069
Na-formate 3.20 g 3.2 47.054
Na-acetate 1.00 g 1 12.19
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 -
Na2CO3 1.00 g 1 9.435
Seven vitamins solution 1.00 ml - -
L-Cysteine HCl x H2O 0.25 g 0.25 1.423
Na2S x 9 H2O 0.25 g 0.25 1.041
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonate, vitamins, cysteine and sulfide, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins (sterilized by filtration), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use adjust pH of complete medium to 8.0 - 8.5.

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.

Seven vitamins solution #

1000
Solution occurs in 78 other media (first defined in Medium 503).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin B12 100 mg 0.1 0.074
p-Aminobenzoic acid 80 mg 0.08 0.583
D-(+)-biotin 20 mg 0.02 0.082
Nicotinic acid 200 mg 0.2 1.625
Calcium pantothenate 100 mg 0.1 0.21
Pyridoxine hydrochloride 300 mg 0.3 1.459
Thiamine-HCl x 2 H2O 200 mg 0.2 0.536
Distilled water 1000 ml - -

Metadata

Last modified: 07.04.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 8.0 - 8.5
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 100 % N2

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

Associated strains

All strains for this medium: 2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na-formate 3.2 47.054
Tryptone 1 14.069
Na-acetate 1 12.19
Na2CO3 1 9.435
K2HPO4 0.5 2.871
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Cultivation metadata from 2 strains BacDive

phyla 1 2 strains
oxygen anaerobe 2 strains
temperature mesophilic 2 strains