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DSMZ

DSMZ 169b . LEADBETTERA MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 169b #

ml
Equipment needed: Autoclave Hungate tubes Gassing station Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 952 ml - -
Solution B 20 ml - -
Solution C 20 ml - -
Solution D 1 ml - -
Solution E 10 ml - -
1 Dissolve ingredients of solution A (except cysteine) and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add cysteine and adjust pH 7.2 with KOH, then distribute under the same gas atmosphere in anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere, solution C is autoclaved under 100% N2 gas atmosphere and solutions D and E are sterilized by filtration under 100% N2 gas atmosphere. To complete the medium appropriate amounts of solutions B, C, D and E are added to the sterile solution A in the sequence as indicated. Final pH of the complete medium should be 7.5.

Solution A #

952
Compound Amount Unit Conc. [g/L] Conc. [mM]
CaCl2 x 2 H2O 0.04 g 0.042 0.286
MgSO4 x 7 H2O 0.50 g 0.525 2.131
Yeast extract
(OXOID) 		4.00 g 4.202 -
D-Glucose 4.00 g 4.202 23.322
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
L-Cysteine HCl x H2O 0.50 g 0.525 2.99
Distilled water 950.00 ml - -

Solution B #

20
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaHCO3 1 g 50 595.188
Distilled water 20 ml - -

Solution C #

20
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4
(0.5 M) 		16.04 ml - -
KH2PO4
(0.5 M) 		3.96 ml - -
1 Combine 16.04 ml of 0.5 M K2HPO4 with 3.96 ml of 0.5 M KH2PO4 stock solution to reach a pH of around 7.4.

Solution D #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Seven vitamins solution 1 ml - -

Solution E #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
DL-Dithiothreitol 0.15 g 15 97.213
Distilled water 10.00 ml - -

Trace element solution SL-10 #

1000
Solution occurs in 267 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.

Selenite-tungstate solution #

1000
Solution occurs in 180 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 - -

Seven vitamins solution #

1000
Solution occurs in 77 other media (first defined in Medium 503).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin B12 100 mg 0.1 0.074
p-Aminobenzoic acid 80 mg 0.08 0.583
D-(+)-biotin 20 mg 0.02 0.082
Nicotinic acid 200 mg 0.2 1.625
Calcium pantothenate 100 mg 0.1 0.21
Pyridoxine hydrochloride 300 mg 0.3 1.459
Thiamine-HCl x 2 H2O 200 mg 0.2 0.536
Distilled water 1000 ml - -

Metadata

Last modified: 25.10.22
Source: DSMZ
Synonyms: TREPONEMA AZOTONUTRICIUM MEDIUM
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.5
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: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
D-Glucose 3.988 22.136
Yeast extract 3.988 -
NaHCO3 0.997 11.868
L-Cysteine HCl x H2O 0.499 2.838
MgSO4 x 7 H2O 0.499 2.023
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature psychrophilic 1 strain