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DSMZ

DSMZ 1030 . SMITHELLA MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 1030 #

ml
Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.0 g 0.978 18.293
MgCl2 x 6 H2O 1.0 g 0.978 4.813
CaCl2 x 2 H2O 0.4 g 0.391 2.662
KH2PO4 0.4 g 0.391 2.876
Wolfe's mineral elixir 1.0 ml - -
Yeast extract 2.0 g 1.957 -
Trypticase peptone
(BD BBL) 		2.0 g 1.957 -
Sodium resazurin
(0.1% w/v) 		0.5 ml 4.9e-4 0.002
Na2CO3 1.0 g 0.978 9.232
Na-crotonate solution (1 M) 20.0 ml - -
2-Mercaptoethanesulfonate
(coenzyme M) 		0.5 g 0.489 3.44
Wolin's vitamin solution (10x) 1.0 ml - -
Na2S x 9 H2O 0.3 g 0.294 1.222
Distilled water 1000.0 ml - -
1 Dissolve ingredients (except carbonate, crotonate, coenzyme M, vitamins and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into Hungate-type tubes or serum vials (e.g., 5 ml medium in Balch-type tubes) and autoclave. Add crotonate, coenzyme M, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of crotonate, coenzyme M and vitamins should be sterilized by filtration. Adjust pH of the complete medium to 7.0 - 7.2.
2 Note: Use 10% (v/v) as inoculum.

Wolfe's mineral elixir #

1000
Solution occurs in 22 other media (first defined in Medium 792).
Compound Amount Unit Conc. [g/L] Conc. [mM]
MgSO4 x 7 H2O 30.00 g 30 121.716
MnSO4 x H2O 5.00 g 5 29.583
NaCl 10.00 g 10 171.116
FeSO4 x 7 H2O 1.00 g 1 3.597
CoCl2 x 6 H2O 1.80 g 1.8 13.863
CaCl2 x 2 H2O 1.00 g 1 6.802
ZnSO4 x 7 H2O 1.80 g 1.8 6.26
CuSO4 x 5 H2O 0.10 g 0.1 0.401
AlK(SO4)2 x 12 H2O 0.18 g 0.18 0.379
H3BO3 0.10 g 0.1 1.617
Na2MoO4 x 2 H2O 0.10 g 0.1 0.413
(NH4)2Ni(SO4)2 x 6 H2O 2.80 g 2.8 7.089
Na2WO4 x 2 H2O 0.10 g 0.1 0.303
Na2SeO4 0.10 g 0.1 0.529
Distilled water 1000.00 ml - -
1 First adjust pH to 1.0 with diluted H2SO4, then add and dissolve the salts.

Na-crotonate solution (1 M) #

1000
Solution occurs in 3 other media (first defined in Medium 870).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Crotonic acid
(ALDRICH 113018) 		86 g 86 998.955
NaOH
(10 N) 		100 ml - -
Distilled water 900 ml - -
1 Dissolve crotonic acid in 800 ml distilled water, add around 100 ml of 10 N NaOH and adjust pH to around 7. Then add water to reach a volume of 1000 ml. Sterilize by filtration under 100% N2 gas atmosphere.

Wolin's vitamin solution (10x) #

1000
Solution occurs in 308 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: 26.11.24
Source: DSMZ
Medium type: Complex medium
Final pH: 7.0 - 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: No strains found

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Yeast extract 1.957 -
Trypticase peptone 1.957 -
Crotonic acid 1.683 19.549
NH4Cl 0.978 18.293
Na2CO3 0.978 9.232
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Cultivation metadata from 0 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain