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DSMZ

DSMZ 1523 . MODIFIED METHANOBACTERIUM MEDIUM

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

Culture medium recipe

Vol.
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Medium for Methanospirillum purgamenti DSM 3595

This strain uses the standard medium without modifications.

Growth conditions: anaerobic 37 °C

Main sol. 1523 #

ml
Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.50 g 0.499 3.667
MgSO4 x 7 H2O 0.40 g 0.399 1.62
NaCl 0.40 g 0.399 6.831
NH4Cl 0.40 g 0.399 7.463
CaCl2 x 2 H2O 0.05 g 0.05 0.339
Trace element solution SL-10 1.00 ml - -
Brain heart infusion
(BD Bacto) 		6.00 g 5.988 -
Proteose peptone
(BD Difco) 		6.00 g 5.988 -
Yeast extract
(OXOID) 		2.00 g 1.996 -
Na-acetate 1.00 g 0.998 12.166
Na-formate 2.00 g 1.996 29.35
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 4.00 g 3.992 47.52
Seven vitamins solution 1.00 ml - -
L-Cysteine HCl x H2O 0.50 g 0.499 2.841
Na2S x 9 H2O 0.50 g 0.499 2.078
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, vitamins, cysteine, and sulfide, then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate and adjust pH to 6.5. Then dispense medium under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add vitamins, cysteine, and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The vitamin solution should be sterilized by filtration. Before use, check the complete medium's pH and adjust it to 6.8 - 7.0, if necessary.
2 After growth has started and the culture is becoming turbid add sterile 80% H2 and 20% CO2 gas mixture to 0.5 - 1 bar overpressure.

Trace element solution SL-10 #

1000
Solution occurs in 266 other media (first defined in Medium 320).
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(25%) 		10.0 ml 2.5 68.568
FeCl2 x 4 H2O 1.5 g 1.5 7.545
ZnCl2 70.0 mg 0.07 0.514
MnCl2 x 4 H2O 100.0 mg 0.1 0.505
H3BO3 6.0 mg 0.006 0.097
CoCl2 x 6 H2O 190.0 mg 0.19 1.463
CuCl2 x 2 H2O 2.0 mg 0.002 0.012
NiCl2 x 6 H2O 24.0 mg 0.024 0.185
Na2MoO4 x 2 H2O 36.0 mg 0.036 0.149
Distilled water 990.0 ml - -
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.

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: 28.02.23
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 % 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: 36
Strains with modifications: 17
Methanospirillum purgamenti DSM 3595

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Brain heart infusion 5.988 -
Proteose peptone 5.988 -
NaHCO3 3.992 47.52
Na-formate 1.996 29.35
Yeast extract 1.996 -
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Cultivation metadata from 36 strains BacDive

pH 4 - 6 1 strain
pH 6 - 8 1 strain
pH 8 - 10 1 strain
phyla 1 35 strains
oxygen anaerobe 33 strains
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