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DSMZ

DSMZ 395a . BROCKIA MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 395a #

ml
Equipment needed: Filter Hungate tubes Gassing station Water bath
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.33 g 0.329 6.157
KH2PO4 0.33 g 0.329 2.42
KCl 0.33 g 0.329 4.418
CaCl2 x 2 H2O 0.44 g 0.439 2.987
MgCl2 x 6 H2O 0.70 g 0.699 3.436
NaCl 0.50 g 0.499 8.539
Trace element solution SL-10 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Sulfur
(powdered) 		10.00 g 9.98 311.195
NaHCO3 0.80 g 0.798 9.504
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.499 2.078
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except sulfur, bicarbonate, vitamins and sulfide), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Adjust pH to 6.2 - 6.4, and dispense medium under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials containing already the appropriate amount of sulfur. Sterilize medium by heating cultivation vessels in a boiling water bath for 2 - 3 hours on each of 3 successive days. After sterilization add bicarbonate from a sterile stock solution prepared under 80% N2 and 20% CO2 gas mixture. Vitamins (sterilized by filtration) and sulfide are added to the medium from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 6.5, if necessary.

Trace element solution SL-10 #

1000
Solution occurs in 266 other media (first defined in Medium 320).
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(25%) 		10.0 ml 2.5 68.568
FeCl2 x 4 H2O 1.5 g 1.5 7.545
ZnCl2 70.0 mg 0.07 0.514
MnCl2 x 4 H2O 100.0 mg 0.1 0.505
H3BO3 6.0 mg 0.006 0.097
CoCl2 x 6 H2O 190.0 mg 0.19 1.463
CuCl2 x 2 H2O 2.0 mg 0.002 0.012
NiCl2 x 6 H2O 24.0 mg 0.024 0.185
Na2MoO4 x 2 H2O 36.0 mg 0.036 0.149
Distilled water 990.0 ml - -
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.

Wolin's vitamin solution (10x) #

1000
Solution occurs in 306 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: 07.11.22
Source: DSMZ
Synonyms: BROCKIA LITHOTROPHICA MEDIUM
Taxonomic range:  Bacteria
Medium type: Defined medium
Final pH: 6.5
Equipment needed: Filter, Gassing station, Hungate tubes, Water bath
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Gas composition

  • 80 % H2
  • 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: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Sulfur 9.98 311.195
NaHCO3 0.798 9.504
MgCl2 x 6 H2O 0.699 3.436
NaCl 0.499 8.539
Na2S x 9 H2O 0.499 2.078
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Cultivation metadata from 1 strains BacDive

pH 4 - 6 1 strain
pH 6 - 8 1 strain
pH 8 - 10 1 strain
phyla 1 1 strain
oxygen anaerobe 1 strain
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