DSMZ 28a . THIORHODOCOCCUS MEDIUM

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Culture medium recipe

Vol.
×
Compound Amount Unit Conc. [g/L] Conc. [mM]
Medium 28 1000.00 ml - -
NaCl 2.00 % 20 342.231
MgCl2 x 6 H2O 0.10 % 1 4.919
Sodium Thiosulfate 0.05 % 0.5 3.162
1 Incubate at 500 to 1000 lux light intensity.
Solution occurs in 8 other media (first defined in Medium 28).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 460 ml - -
1 Aliquot Solution A into 100 mL screw-cap bottles, filled with 46 mL each. Bubble with N2/CO2 and autoclave at 121°C for 15 min (as decribed below).
2 Prepare the following solutions (resazurin, bicarbonate and Pfennig's heterotrophic salts) and sterilize as given below.
Resazurin solution 450 ml - -
Bicarbonate solution 50 ml - -
Pfennig's heterotrophic salts solution 26 ml - -
3 Add bicarbonate solution and Pfennig's heterotrophic salts to the resazurin (complete volumina, i.e. 50 mL bicarbonate solution and 26 mL Pfennig's heterotrophic salts solution). Bubble with CO2 in an ice bath under sterile conditions.
4 Fill 50 ml of this mixture to each bottle of solution A (46 mL + 50 mL).
5 Before use, add 4 ml sulfide solution (1.5%) and 0.1 ml Vitamin B12 solution to each 100 mL bottle.
Sulfide solution, 1.5% 40 ml - -
Vitamin B12 solution 1 ml - -
6 Adjust the pH with filter-sterilised 1M Na2CO3 to 7.1-7.3.
7 If needed, aliquot into sterile, N2 gassed screw-cap tubes under N2 gas.
8 During the first 24 h, the iron of the medium precipitates in the form of black flocks. No other sediment should arise in the otherwise clear medium.
9 Feed the actively growing culture periodically with neutralized 3% solution of sodium sulfide (use 1 -3 mL/100 mL depending on strain and cultivation stage) to replenish sulfide and with other supplement solutions (see Ref. 3365).
Neutralized sulfide solution 3% (w/v) 10 ml - -
Solution occurs in 13 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: 22.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.1 - 7.3
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Associated strains

All strains for this medium: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 20 342.231
NaHCO3 1.5 17.856
MgCl2 x 6 H2O 1 4.919
Na2S x 9 H2O 0.593 2.467
MgSO4 x 7 H2O 0.5 2.029
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Cultivation metadata from 1 strains BacDive

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