DSMZ 1267a . MARIPROFUNDUS (ES) MEDIUM

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Culture medium recipe

Vol.
×
Equipment needed: Gassing station Filter Autoclave Hungate tubes
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 901 ml - -
Solution B 10 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 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 bottom layer; the pipette tip is drawn upward as the inoculum is dispensed.
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 27.50 g 30.522 522.273
MgCl2 x 6 H2O 5.38 g 5.971 29.371
MgSO4 x 7 H2O 6.78 g 7.525 30.53
KCl 0.72 g 0.799 10.719
CaCl2 x 2 H2O 1.40 g 1.554 10.569
NH4Cl 1.00 g 1.11 20.749
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 - -
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Wolin's vitamin solution 10 ml - -
1 Sterilize by filtration and store at 4°C in the dark.
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ferrous sulfide sludge 100 ml - -
Solution occurs in 23 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 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 - -
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: 11.04.22
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 15.232 54.79
Na2S x 9 H2O 12.166 50.654
MgCl2 x 6 H2O 5.321 26.175
Agarose 1.484 2.353
NH4Cl 0.989 18.492
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Cultivation metadata from 0 strains BacDive

phyla 1 1 strain
oxygen microaerophile 1 strain
temperature mesophilic 1 strain