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DSMZ

DSMZ 377 . PYROCOCCUS MEDIUM

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

Culture medium recipe

Vol.
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Medium for Pyrococcus kukulkanii DSM 101590

This strain uses the standard medium without modifications.

Growth conditions: anaerobic 100 °C

Main sol. 377 #

ml
Equipment needed: Hungate tubes Gassing station Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 13.85 g 13.85 236.995
MgSO4 x 7 H2O 3.50 g 3.5 14.2
MgCl2 x 6 H2O 2.75 g 2.75 13.527
KCl 0.33 g 0.33 4.427
NaBr 0.05 g 0.05 0.486
H3BO3 15.00 mg 0.015 0.243
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 0.75 5.102
KH2PO4 0.50 g 0.5 3.674
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 30 935.454
Peptone
(BD Bacto) 		5.00 g 5 -
Yeast extract
(OXOID) 		1.00 g 1 -
Neutralized sulfide solution 3% (w/v) 15.00 ml - -
Distilled water 960.00 ml - -
1 Dissolve ingredients except for sulfur, peptone, yeast extract, 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 and yeast extract from sterile stock solutions prepared under 100% N2 gas atmosphere and sulfide from a neutralized stock solution.

Modified Wolin's mineral solution #

1000
Solution occurs in 167 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.

Neutralized sulfide solution 3% (w/v) #

100
Solution occurs in 19 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, Gassing station, Hungate tubes
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Gas composition

  • 100 % N2

Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .

Associated strains

All strains for this medium: 10
Pyrococcus kukulkanii DSM 101590

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Sulfur 30 935.454
Peptone 5 -
MgCl2 x 6 H2O 2.75 13.527
Yeast extract 1 -
KH2PO4 0.5 3.674
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Cultivation metadata from 10 strains BacDive

pH 2 - 4 1 strain
pH 6 - 8 1 strain
pH 8 - 10 1 strain
phyla 3 10 strains
oxygen anaerobe 8 strains
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