DSMZ 1267b . GALLIONELLA (ES) MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 901 ml - -
Solution B 1 ml - -
Solution C 100 ml - -
1 Preparation of the bottom layer: Mix 1 volume of solution A (except NaHCO3 and agarose) with 1 volume of solution C and add 1% (w/v) agarose type 1, low EEO. After autoclaving, aseptically fill 1.2 ml of the suspension in sterile screw capped tubes (10 ml total volume). The bottom layer solidifies in approx. 30 min.
2 Preparation of the top layer: Add bicarbonate and low melt agarose to solution A and autoclave. Let the sterile solution cool to 40°C and add 10.00 ml/l of solution B. Sparge solution with sterile 100% CO2 gas until a pH of 6.1 - 6.4 is reached. Then, aseptically pipette aliquots of 6.0 ml over the bottom layer of each tube and let medium equilibrate for at least three hours, but not longer than 12 hours.
3 Inoculation: Inoculate the semisolid top layer with a pipette that is inserted just above the FeS layer; the pipette tip is drawn upward as the inoculum is dispensed.
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.00 g 1.11 20.749
MgSO4 x 7 H2O 0.20 g 0.222 0.901
CaCl2 x 2 H2O 0.10 g 0.111 0.755
K2HPO4 0.05 g 0.055 0.319
Wolfe's mineral elixir 1.00 ml - -
NaHCO3
(for top layer)
0.50 g 0.555 6.606
Agarose
(for top layer, low melt)
1.50 g 1.665 2.64
Distilled water 900.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Wolin's vitamin solution (10x) 1 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ferrous sulfide sludge 100 ml - -
Solution occurs in 21 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.
Solution occurs in 299 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 - -
Solution occurs in 3 other media (first defined in Medium 1267).
Compound Amount Unit Conc. [g/L] Conc. [mM]
FeSO4 x 7 H2O 15.4 g 154 553.927
Na2S x 9 H2O 12.3 g 123 512.112
Distilled water 100.0 ml - -
1 Heat distilled water to 50°C in a 250 ml beaker with a stir bar present. While rapidly stirring the water, add the ferrous sulfate followed immediately by the sodium sulfide. The formed black FeS sludge is decanted into a glass bottle that can be stoppered. The FeS is allowed to settle for several hours and then the overlying water is decanted and replaced. This procedure is repeated at least five times to wash the FeS. After washing, the pH of the FeS solution should be close to neutrality. The FeS suspension can be kept in closed bottles or tubes under a nitrogen atmosphere for at least three months.

Metadata

Last modified: 19.09.22
Source: DSMZ
Medium type: Defined medium
Final pH: 6.1 - 6.4
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Associated strains

All strains for this medium: No strains found

Molecular composition

Compound Conc. [g/L] Conc. [mM]
FeSO4 x 7 H2O 15.369 55.282
Na2S x 9 H2O 12.275 51.109
Agarose 1.497 2.374
NH4Cl 0.998 18.658
NaHCO3 0.499 5.94
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