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DSMZ

DSMZ 377a . PALAEOCOCCUS MEDIUM

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

Culture medium recipe

Vol.
×
Medium for Lebetimonas natsushimae DSM 104102

* Modification: Add 2 bar overpressuere of sterile 80% H2 and 20% CO2 gas mixture after inoculation.

Growth conditions: anaerobic 55 °C

Main sol. 377a #

ml
Equipment needed: Hungate tubes Filter Gassing station Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 30.00 g 29.97 512.834
MgSO4 x 7 H2O 3.50 g 3.497 14.186
MgCl2 x 6 H2O 2.75 g 2.747 13.513
KCl 0.33 g 0.33 4.422
NaBr 0.05 g 0.05 0.485
H3BO3 15.00 mg 0.015 0.242
SrCl2 x 6 H2O
(0.1% w/v) 		7.00 ml 0.007 0.026
(NH4)2SO4 10.00 mg 0.01 0.076
Citric acid
(0.1% w/v) 		5.00 ml 0.005 0.026
KI
(0.01% w/v) 		0.50 ml 5.0e-5 1.9e-4
CaCl2 x 2 H2O 0.75 g 0.749 5.096
KH2PO4 0.50 g 0.5 3.671
NiCl2 x 6 H2O
(0.1% w/v) 		2.00 ml 0.002 0.015
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Sulfur
(powdered) 		30.00 g 29.97 934.519
Peptone
(BD Bacto) 		5.00 g 4.995 -
Yeast extract
(OXOID) 		1.00 g 0.999 -
Wolin's vitamin solution (10x) 1.00 ml - -
Neutralized sulfide solution 3% (w/v) 15.00 ml - -
Distilled water 960.00 ml - -
1 Dissolve ingredients except sulfur, peptone, yeast extract, vitamins, and sulfide, adjust the pH to 6.5 and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Distribute medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials that already contain the appropriate amount of sulfur. Sterilize the medium by autoclaving at 110°C for 20 min. After sterilization add peptone, yeast extract, and vitamins from sterile stock solutions prepared under 100% N2 gas atmosphere and sulfide from a neutralized stock solution. Vitamins should be sterilized by filtration.

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 - -

Neutralized sulfide solution 3% (w/v) #

100
Solution occurs in 18 other media (first defined in Medium 28).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 3 g 30 124.905
Distilled water 100 ml - -
1 The sulfide solution is prepared in a 250 ml screw-capped bottle with a butyl rubber septum and a magnetic stirrer. The solution is bubbled with nitrogen gas, closed and autoclaved for 15 min. at 121°C. After cooling to room temperature the pH is adjusted to about 7.0 by adding of sterile 2 M H2SO4 drop-wise with a syringe without opening the bottle.

Metadata

Last modified: 15.12.22
Source: DSMZ
Taxonomic range:  Archaea
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 6.5
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 53 % H2
  • 33 % N2
  • 13 % 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: 6
Strains with modifications: 1
Lebetimonas natsushimae DSM 104102

  33% N2, 53% H2, 13% CO2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Sulfur 29.97 934.519
Peptone 4.995 -
MgCl2 x 6 H2O 2.747 13.513
Yeast extract 0.999 -
KH2PO4 0.5 3.671
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Cultivation metadata from 6 strains BacDive

pH 4 - 6 1 strain
pH 6 - 8 1 strain
phyla 3 5 strains
oxygen anaerobe 4 strains
temperature hyperthermophilic 3 strains
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