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DSMZ

DSMZ 1597 . METHANOSALSUM NATRONOPHILUM MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 1597 #

ml
Equipment needed: Gassing station Autoclave Hungate tubes Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 750 ml - -
Solution B 250 ml - -
Solution C 1 ml - -
Solution D 1 ml - -
Solution E 5 ml - -
Solution F 3 ml - -
Solution G 5 ml - -
Solution H 10 ml - -
Solution I 1 ml - -
Solution J 10 ml - -
1 Sterilize solutions A and B in closed thick-walled screw-top bottles (e.g., SCHOTT) for 20 min at 120°C. The pH of solution A after sterilization should be 10. There is some precipitate forming that settles at the bottom after 3 - 4 days. It is best to remove precipitates by decantation before using solution A for medium preparation.
2 Combine solution A with solution B and sparge medium with 100% N2 gas for at least 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solutions C to J are sterilized separately under 100% N2 gas. Vitamins and coenzyme M should be sterilized by filtration. To complete the medium appropriate amounts of solutions C to J are added to the combined sterile solutions A and B in the sequence as indicated. Final pH of the medium should be 9.5.
3 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized) may stimulate growth at the beginning. For transfers use 5 - 10% inoculum.

Solution A #

750
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 71.25 g 95 896.328
NaHCO3 11.25 g 15 178.557
NaCl 4.00 g 5.333 91.262
K2HPO4 0.75 g 1 5.741
Distilled water 750.00 ml - -

Solution B #

250
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 30.00 g 120 2053.39
K2HPO4 0.25 g 1 5.741
KCl 0.25 g 1 13.414
Distilled water 250.00 ml - -

Solution C #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Trace elements solution (Pfennig & Lippert,1966) 1 ml - -

Solution D #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Selenite-tungstate solution 1 ml - -

Solution E #

5
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.20 g 40 747.803
MgSO4 x 7 H2O 0.25 g 50 202.86
Distilled water 5.00 ml - -

Solution F #

3
Compound Amount Unit Conc. [g/L] Conc. [mM]
Methanol 3 ml 792 24717.6

Solution G #

5
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract 0.02 g 4 -
Distilled water 5.00 ml - -

Solution H #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
2-Mercaptoethanesulfonic acid
(coenzyme M) 		0.15 g 15 105.485
Distilled water 10.00 ml - -

Solution I #

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

Solution J #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 0.25 g 25 104.088
Distilled water 10.00 ml - -

Trace elements solution (Pfennig & Lippert,1966) #

1000
Solution occurs in 28 other media (first defined in Medium 1369).
Compound Amount Unit Conc. [g/L] Conc. [mM]
EDTA 5.00 g 5 17.109
FeSO4 x 7 H2O 2.20 g 2.2 7.913
ZnSO4 x 7 H2O 0.10 g 0.1 0.348
MnCl2 x 4 H2O 0.03 g 0.03 0.152
H3BO3 0.03 g 0.03 0.485
CoCl2 x 6 H2O 0.20 g 0.2 1.54
CuCl2 x 2 H2O 0.03 g 0.03 0.176
NiCl2 x 6 H2O 0.03 g 0.03 0.231
Na2MoO4 x 2 H2O 0.03 g 0.03 0.124
Distilled water 1000.00 ml - -
1 pH 3.0-4.0

Selenite-tungstate solution #

1000
Solution occurs in 178 other media (first defined in Medium 385).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH 0.5 g 0.5 12.501
Na2SeO3 x 5 H2O 3.0 mg 0.003 0.011
Na2WO4 x 2 H2O 4.0 mg 0.004 0.012
Distilled water 1000.0 ml - -

Wolin's vitamin solution (10x) #

1000
Solution occurs in 305 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: 20.09.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 9.5
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: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na2CO3 68.774 648.886
NaCl 28.958 495.509
NaHCO3 10.859 129.264
Methanol 2.293 71.576
KCl 0.241 3.237
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain