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DSMZ

DSMZ 924b . METHANOSARCINA MB MEDIUM (1 g/l NaCl)

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

Culture medium recipe

Vol.
×
Medium for Methanosarcina mazei DSM 27785

* Modification: Omit formate and acetate and supplement medium with 4.00 ml/l methanol added from a sterile anoxic stock solution.

Growth conditions: 2-3 days anaerobic 37 °C

Main sol. 924b #

ml
Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 1.00 g 0.989 16.925
MgCl2 x 6 H2O 1.00 g 0.989 4.865
KCl 0.50 g 0.495 6.634
CaCl2 x 2 H2O 0.40 g 0.396 2.691
K2HPO4 0.40 g 0.396 2.272
NH4Cl 1.00 g 0.989 18.492
Modified Wolin's mineral solution 10.00 ml - -
Yeast extract
(OXOID) 		2.00 g 1.978 -
Trypticase peptone
(BD BBL) 		2.00 g 1.978 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
NaHCO3 4.00 g 3.956 47.097
Na-formate* 6.80 g 6.726 98.902
Na-acetate* 1.60 g 1.583 19.292
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.25 g 0.247 1.408
Na2S x 9 H2O 0.25 g 0.247 1.03
Methanol* 4.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, formate, acetate 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 Hungate-type tubes or serum vials and autoclave. Add formate, acetate, 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.

Modified Wolin's mineral solution #

1000
Solution occurs in 166 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.

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: 08.04.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 7.2
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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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: 3
Strains with modifications: 2
Methanosarcina mazei DSM 27785

  4 ml Methanol
  Na-formate
  Na-acetate

Molecular composition
* modified beta

Compound Conc. [g/L] Conc. [mM]
NaHCO3 3.956 47.097
Methanol 3.134 97.795
Yeast extract 1.978 -
Trypticase peptone 1.978 -
NH4Cl 0.989 18.492
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Cultivation metadata from 3 strains BacDive

phyla 1 3 strains
oxygen anaerobe 3 strains
temperature mesophilic 3 strains