DSMZ 559 . TMBS4 MEDIUM

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Culture medium recipe

Vol.
×
Equipment needed: Gassing station Autoclave Hungate tubes Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 932 ml - -
Solution B 30 ml - -
Solution C 10 ml - -
Solution D 2 ml - -
Solution E 10 ml - -
Solution F 10 ml - -
Solution G 10 ml - -
1 Boil solution A for 3 min., then cool to room temperature under 100% N2 gas atmosphere. Dispense the medium under same gas atmosphere in culture vessels and autoclave. Autoclave separately solutions E and F under 100% N2 and solution B under 80% N2 and 20% CO2 gas atmosphere. Solutions C, D, and G are prepared under N2 gas atmosphere and sterilized by filtration.
2 Solutions B to G are added to the sterile, cooled solution A in the sequence as indicated. The final pH of the medium should be 7.2-7.4. After inoculation add 25 mg sodium dithionite per liter medium from a 2.5% (w/v) solution, freshly prepared under N2 and filter-sterilized.
Equipment needed: Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 1.00 g 1.073 18.36
MgCl2 x 6 H2O 0.40 g 0.429 2.111
CaCl2 x 2 H2O 0.15 g 0.161 1.095
KCl 0.50 g 0.536 7.196
NH4Cl 0.25 g 0.268 5.015
KH2PO4 0.20 g 0.215 1.577
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Resazurin 0.50 mg 5.4e-4 0.002
Distilled water 930.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaHCO3 , 5% w/v solution 30 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin solution 10 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-thiosulfate , 5% w/v solution 2 ml 1667 10543.3
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract , 10% w/v solution 10 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Syringate solution 6.0 %(w/v) 10 ml - -
Equipment needed: Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Dithiothreitol , 100 mM solution 10 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 2 mg 0.002 0.008
Folic acid 2 mg 0.002 0.005
Pyridoxine hydrochloride 10 mg 0.01 0.049
Thiamine HCl 5 mg 0.005 0.015
Riboflavin 5 mg 0.005 0.013
Nicotinic acid 5 mg 0.005 0.041
Calcium D-(+)-pantothenate 5 mg 0.005 -
Vitamin B12 5 mg 0.005 0.004
p-Aminobenzoic acid 5 mg 0.005 0.036
(DL)-alpha-Lipoic acid 5 mg 0.005 0.024
Distilled water 1000 ml - -
1 Adjust final pH of solution to 7.2 using 50 mM sodium phosphate buffer.
Compound Amount Unit Conc. [g/L] Conc. [mM]
Syringic acid 6 %(w/v) 60 -
1 Dissolve syringic acid in distilled water and neutralize solution with 2 N NaOH until a pH of 7.5 is reached. The final concentration of syringate in the solution should be 6% w/v.
Solution occurs in 263 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.
Solution occurs in 176 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 - -

Metadata

Last modified: 28.03.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.2 - 7.4
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]
Na-thiosulfate 3.321 21.003
NaCl 0.996 17.043
Syringic acid 0.598 -
KCl 0.498 6.68
MgCl2 x 6 H2O 0.398 1.96
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain