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DSMZ

DSMZ 334 . METHANOTHRIX MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 334 #

ml
Equipment needed: Gassing station Serum bottle Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 951 ml - -
Solution B 25 ml - -
Solution C 10 ml - -
Solution D 10 ml - -
Solution E 1 ml - -
Solution F 10 ml - -
1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, dispense under same gas atmosphere into anoxic serum vials (e.g., 20 ml medium in 50 ml bottles) and autoclave. Prior to inoculation complete the medium by adding appropriate amounts of sterile solutions C, D, E and F prepared under 100% N2 gas and solution B prepared under 80% N2 and 20% CO2 gas atmosphere. Vitamins are sterilized by filtration. Adjust the pH of the complete medium to 7.2.
2 Note: Use 20% (v/v) inoculum.

Solution A #

951
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.22 g 0.231 1.7
Na2HPO4 x 2 H2O 0.86 g 0.904 6.37
Na-acetate 6.80 g 7.15 87.164
Na2-EDTA
(0.1% w/v) 		1.00 ml 0.001 0.003
Trace elements solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.3e-4 0.002
Distilled water 940.00 ml - -

Solution B #

25
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1.25 g 50 471.752
Distilled water 25.00 ml - -

Solution C #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 0.30 g 30 513.347
CaCl2 x 2 H2O 0.11 g 11 74.822
MgCl2 x 6 H2O 0.10 g 10 49.188
Distilled water 10.00 ml - -

Solution D #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.3 g 30 560.852
Distilled water 10.0 ml - -

Solution E #

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

Solution F #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 0.5 g 50 208.175
Distilled water 10.0 ml - -

Trace elements solution #

1000
Solution occurs in 1 other media (first defined in Medium 334).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid
(NTA) 		12.80 g 12.8 66.967
FeCl3 x 6 H2O 1.35 g 1.35 4.995
MnCl2 x 4 H2O 0.10 g 0.1 0.505
CoCl2 x 6 H2O 0.03 g 0.03 0.231
CaCl2 x 2 H2O 0.10 g 0.1 0.68
ZnCl2 0.10 g 0.1 0.734
CuCl2 0.03 g 0.03 0.223
H3BO3 0.01 g 0.01 0.162
Na2MoO4 x 2 H2O 0.03 g 0.03 0.124
NiCl2 x 6 H2O 0.12 g 0.12 0.926
NaCl 1.00 g 1 17.112
Na2SeO3 x 5 H2O 0.03 g 0.03 0.114
Distilled water 1000.00 ml - -
1 First dissolve NTA in 200 ml of distilled water and adjust pH to 6.5 with KOH, then dissolve mineral salts. Finally adjust pH to 6.5 with KOH and make up to 1000.00 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: 21.09.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Defined medium
Final pH: 7.2
Equipment needed: Autoclave, Filter, Gassing station, Serum bottle
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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: 1
Strains with modifications: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na-acetate 6.753 82.317
Na2CO3 1.241 11.712
Na2HPO4 x 2 H2O 0.854 6.016
Na2S x 9 H2O 0.497 2.067
NH4Cl 0.298 5.57
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Cultivation metadata from 1 strains BacDive

pH 6 - 8 1 strain
pH 8 - 10 1 strain
phyla 1 3 strains
oxygen anaerobe 3 strains
temperature mesophilic 1 strain