DSMZ 924 . METHANOCALCULUS MB MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Autoclave Filter Gassing station Serum bottle
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 10.00 g 10 171.116
MgCl2 x 6 H2O 1.00 g 1 4.919
KCl 0.50 g 0.5 6.707
CaCl2 x 2 H2O 0.40 g 0.4 2.721
K2HPO4 0.40 g 0.4 2.297
NH4Cl 1.00 g 1 18.695
Modified Wolin's mineral solution 10.00 ml - -
Yeast extract 2.00 g 2 -
Trypticase peptone
(BD BBL)
2.00 g 2 -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
NaHCO3 4.00 g 4 47.615
Na-acetate 1.60 g 1.6 19.504
Na-formate 6.80 g 6.8 99.989
Wolin's vitamin solution 10.00 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 bicarbonate, acetate, formate, vitamins and reducing agents), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add solid bicarbonate, adjust pH to 6.8 - 7.0, dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic serum bottles and autoclave. Add acetate, formate, vitamins (sterilized by filtration), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The pH of the complete medium should be 7.2.
Solution occurs in 170 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.
Solution occurs in 357 other media (first defined in Medium 141).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 2.0 mg 0.002 0.008
Folic acid 2.0 mg 0.002 0.005
Pyridoxine hydrochloride 10.0 mg 0.01 0.049
Thiamine HCl 5.0 mg 0.005 0.015
Riboflavin 5.0 mg 0.005 0.013
Nicotinic acid 5.0 mg 0.005 0.041
Calcium D-(+)-pantothenate 5.0 mg 0.005 -
Vitamin B12 0.1 mg 1.0e-4 7.4e-5
p-Aminobenzoic acid 5.0 mg 0.005 0.036
(DL)-alpha-Lipoic acid 5.0 mg 0.005 0.024
Distilled water 1000.0 ml - -

Metadata

Last modified: 08.04.22
Source: DSMZ
Taxonomic range:  Archaea
Taxonomic range:  Bacteria
Medium type: Complex 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: 6
Strains with modifications: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na-formate 6.8 99.989
NaHCO3 4 47.615
Trypticase peptone 2 -
Yeast extract 2 -
Na-acetate 1.6 19.504
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Cultivation metadata from 6 strains BacDive

phyla 1 6 strains
oxygen anaerobe 5 strains
temperature mesophilic 6 strains