DSMZ 28 . PFENNIG'S MEDIUM I

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Culture medium recipe

Vol.
×
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 4 other media (first defined in Medium 28).
Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
CaCl2 x 2 H2O 0.25 g 0.543 3.697
Yeast extract 0.25 g 0.543 -
Distilled water 460.00 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.
Solution occurs in 4 other media (first defined in Medium 28).
Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 2 g 14.815 61.682
Distilled water 135 ml - -
1 Prepare in a screw-cap bottle, bubble with N2 to replace air, close tightly and autoclave.
Solution occurs in 4 other media (first defined in Medium 28).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaHCO3 1.5 g 30 357.113
H2O 50.0 ml - -
1 Bubble with CO2 and filter sterilize into sterile, gas-tight, 100 ml screw-cap bottle.
Solution occurs in 4 other media (first defined in Medium 28).
Equipment needed: Autoclave Ice bath
Compound Amount Unit Conc. [g/L] Conc. [mM]
Resazurin
(0,1%)
0.5 ml 0.005 0.022
Distilled water 450.0 ml - -
1 Autoclave in a cotton-stoppered Erlenmeyer flask with an outlet tube for medium, connected to a glass outlet at the bottom of the vessel and has, at the other end, a silicon rubber tube with a pinch cock and a bell for aseptic dispensing of the medium into bottles.
2 Cool to room temperature under an atmosphere of N2/CO2 in an ice bath.
Solution occurs in 4 other media (first defined in Medium 28).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ammonium chloride 0.35 g 13.462 251.665
Ammonium acetate 0.25 g 9.615 124.746
Pyruvic acid sodium salt 0.25 g 9.615 87.381
Dextrose 0.25 g 9.615 48.521
MgSO4 x 7 H2O 0.50 g 19.231 78.023
KCl 0.35 g 13.462 180.571
KH2PO4 0.35 g 13.462 98.92
Trace element solution SL-12 B 1.00 ml - -
Distilled water 25.00 ml - -
1 Filter sterilize into sterile, gas-tight, 100 ml screw-cap bottle.
Solution occurs in 5 other media (first defined in Medium 28).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin B12 0.01 g 0.1 0.074
Distilled water 100.00 ml - -
1 Filter sterilized
Solution occurs in 3 other media (first defined in Medium 28).
Equipment needed: Ice bath Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2-EDTA 3.0 g 3 8.923
FeSO4 x 7 H2O 1.1 g 1.1 3.957
CoCl2 x 6 H2O 190.0 mg 0.19 1.463
MnCl2 x 2 H2O 50.0 mg 0.05 -
ZnCl2 42.0 mg 0.042 0.308
NiCl2 x 6 H2O 24.0 mg 0.024 0.185
Na2MoO4 x 2 H2O 18.0 mg 0.018 0.074
H3BO3 300.0 mg 0.3 4.851
CuCl2 x 2 H2O 2.0 mg 0.002 0.012
Distilled water 1000.0 ml - -
1 Adjust pH to 6.0.
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: 24.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.1 - 7.3
Equipment needed: Autoclave, Filter, Ice bath
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Associated strains

All strains for this medium: 59
Strains with modifications: 29

Light

2 strains need light for growth. Select a strain for more details.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaHCO3 1.5 17.856
Na2S x 9 H2O 0.593 2.467
MgSO4 x 7 H2O 0.5 2.029
Ammonium chloride 0.35 6.543
KCl 0.35 4.695
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Cultivation metadata from 59 strains BacDive

pH 6 - 8 3 strains
phyla 1 52 strains
oxygen anaerobe 49 strains
oxygen aerobe 1 strain
oxygen obligate anaerobe 1 strain
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