DSMZ 29 . PFENNIG'S MEDIUM II

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Culture medium recipe

Vol.
×
Solution occurs in 1 other media (first defined in Medium 29).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 460 ml - -
Solution B 40 ml - -
Solution C 50 ml - -
Solution D 450 ml - -
Solution E 26 ml - -
Solution F 10 ml - -
1 Mix solution D, C and E. Bubble with CO2 in an ice bath under sterile conditions.
2 Fill 50 ml in each bottle of solution A. Before using add 4 ml solution B and 0.1 ml solution F.
3 Adjust the pH with filter-sterilised 1M Na2CO3 to 6.8 -7.1.
4 Fill in sterile, N2 gassed screw-cap tubes under N2 gas.
5 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. Feed periodically with neutralized 3% solution of sodium sulfide to replenish sulfide and with other supplement solutions (see Ref. 3365).
Solution occurs in 2 other media (first defined in Medium 29).
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 Fill 10x 46ml in 100 ml screw-cap bottles. Bubble with N2/CO2 and autoclave 121°C 15 min.
2 (For marine or estuarine isolates add 100.0 g NaCl to this solution and increase the MgSO4 x 7 H2O to 15.0 g).
Solution occurs in 2 other media (first defined in Medium 29).
Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Distilled water 135 ml - -
1 Prepare in a screw-cap bottle, bubble with N2 to replace air, close tightly and autoclave.
Solution occurs in 2 other media (first defined in Medium 29).
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 2 other media (first defined in Medium 29).
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 2 other media (first defined in Medium 29).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ammonium chloride 0.35 g 14 261.731
Ammonium acetate 0.25 g 10 129.735
Pyruvic acid sodium salt 0.25 g 10 90.876
Dextrose 0.25 g 10 50.462
MgSO4 x 7 H2O 0.50 g 20 81.144
KCl 0.35 g 14 187.793
KH2PO4 0.35 g 14 102.877
Trace element solution SL-10 B 1.00 ml - -
Distilled water 25.00 ml - -
1 Filter sterilize into sterile, gas-tight, 100 ml screw-cap bottle.
Solution occurs in 2 other media (first defined in Medium 29).
Equipment needed: Filter Ice bath
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 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.
Solution occurs in 1 other media (first defined in Medium ).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Distilled water 1000.0 ml - -
HCl
(25%)
7.7 ml 1.925 52.798
FeSO4 x 7 H2O 1.5 g 1.5 5.395
ZnCl2 70.0 mg 0.07 0.514
MnCl2 x 4 H2O 100.0 mg 0.1 0.505
H3BO3 300.0 mg 0.3 4.851
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

Metadata

Last modified: 24.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 6.8 - 7.1
Equipment needed: Autoclave, Filter, Ice bath
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Associated strains

All strains for this medium: 11
Strains with modifications: 1

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 0.52 2.11
Ammonium chloride 0.364 6.805
KCl 0.364 4.883
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Cultivation metadata from 11 strains BacDive

phyla 1 11 strains
nutrition phototroph 3 strains
oxygen anaerobe 11 strains
oxygen obligate anaerobe 3 strains
temperature mesophilic 10 strains
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