DSMZ 1597b . NATRANAEROBIALES MEDIUM

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Culture medium recipe

Vol.
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Equipment needed: Hungate tubes Gassing station Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 500 ml - -
Solution B 500 ml - -
Solution C 1 ml - -
Solution D 1 ml - -
Solution E 5 ml - -
Solution F 10 ml - -
Solution G 5 ml - -
Solution H 10 ml - -
Solution I 10 ml - -
1 Sterilize solutions A and B in closed thick-walled screw-top bottles (e.g., SCHOTT) for 20 min at 120°C. The pH of solution A after sterilization should be 10. There is some precipitate forming that settles at the bottom after 3 - 4 days. It is best to remove precipitates by decantation before using solution A for medium preparation.
2 Combine solution A with solution B and sparge medium with 100% N2 gas for at least 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solutions C to I are sterilized separately under 100% N2 gas. Vitamins and pyruvate should be sterilized by filtration. To complete the medium appropriate amounts of solutions C to I are added to the combined sterile solutions A and B in the sequence as indicated. Final pH of the medium should be 9.5.
3 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized) may stimulate growth at the beginning. For transfers use 5 - 10% inoculum.
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 92.5 g 185 1745.48
NaHCO3 17.5 g 35 416.632
NaCl 8.0 g 16 273.785
K2HPO4 0.5 g 1 5.741
Distilled water 500.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 120.00 g 240 4106.78
K2HPO4 1.25 g 2.5 14.353
KCl 2.50 g 5 67.069
Distilled water 500.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Trace elements solution (Pfennig & Lippert,1966) 1 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Selenite-tungstate solution 1 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.20 g 40 747.803
MgSO4 x 7 H2O 0.25 g 50 202.86
Distilled water 5.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-pyruvate 2.2 g 220 1999.2
Distilled water 10.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract 0.5 g 100 -
Distilled water 5.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Wolin's vitamin solution 10 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 0.25 g 25 104.088
Distilled water 10.00 ml - -
Solution occurs in 27 other media (first defined in Medium 1369).
Compound Amount Unit Conc. [g/L] Conc. [mM]
EDTA 5.00 g 5 17.109
FeSO4 x 7 H2O 2.20 g 2.2 7.913
ZnSO4 x 7 H2O 0.10 g 0.1 0.348
MnCl2 x 4 H2O 0.03 g 0.03 0.152
H3BO3 0.03 g 0.03 0.485
CoCl2 x 6 H2O 0.20 g 0.2 1.54
CuCl2 x 2 H2O 0.03 g 0.03 0.176
NiCl2 x 6 H2O 0.03 g 0.03 0.231
Na2MoO4 x 2 H2O 0.03 g 0.03 0.124
Distilled water 1000.00 ml - -
1 pH 3.0-4.0
Solution occurs in 174 other media (first defined in Medium 385).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH 0.5 g 0.5 12.501
Na2SeO3 x 5 H2O 3.0 mg 0.003 0.011
Na2WO4 x 2 H2O 4.0 mg 0.004 0.012
Distilled water 1000.0 ml - -
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 - -

Metadata

Last modified: 22.02.22
Source: DSMZ
Medium type: Complex medium
Final pH: 9.5
Equipment needed: Autoclave, 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]
NaCl 115.163 1970.62
Na2CO3 88.772 837.562
NaHCO3 16.795 199.919
KCl 2.399 32.183
Na-pyruvate 2.111 19.186
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