DSMZ 710 . THERMOANAEROBACTER (KoKo) MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Tryptone
(BD Bacto)
1.00 g 1 14.069
Peptone
(meat)
1.00 g 1 -
Yeast extract
(BD Bacto)
1.00 g 1 -
K2HPO4 1.60 g 1.6 9.186
NaH2PO4 x 2 H2O 1.00 g 1 8.335
NH4Cl 0.50 g 0.5 9.348
MgSO4 x 6 H2O 0.16 g 0.16 1.329
Trace element solution SL-11 1.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
CaCl2 x 2 H2O 0.06 g 0.06 0.408
NaHCO3 1.00 g 1 11.904
D-Glucose 5.00 g 5 27.753
Wolin's vitamin solution 10.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.3 1.708
Na2S x 9 H2O 0.30 g 0.3 1.249
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except calcium chloride, bicarbonate, glucose, vitamins, cysteine and sulfide), adjust pH to 7.0 and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add calcium chloride, glucose, cysteine and sulfide from sterile anoxic stock solutions autoclaved under 100% N2 gas atmosphere. Add bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Vitamins are prepared under 100% N2 gas and sterilized by filtration. The pH of the complete medium should be at 7.0.
2 Note: Supplementation of medium with 10.00 g/l MOPS buffer (pH 6.9 - 7.0; 10% (w/v) anoxic stock solution) may enhance the buffer capacity of the medium.
Solution occurs in 22 other media (first defined in Medium 722).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2-EDTA x 2 H2O 5.2 g 5.2 15.467
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 1000.0 ml - -
1 Dissolve EDTA in 800 ml distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add other compounds. Finally adjust pH to 6.0 and bring volume to 1000 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: 06.04.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.0
Equipment needed: Autoclave, Filter, 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: 2
Strains with modifications: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
D-Glucose 5 27.753
K2HPO4 1.6 9.186
Tryptone 1 14.069
NaHCO3 1 11.904
NaH2PO4 x 2 H2O 1 8.335
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Cultivation metadata from 2 strains BacDive

phyla 1 2 strains
oxygen anaerobe 2 strains
temperature thermophilic 2 strains