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DSMZ

DSMZ 29 . PFENNIG'S MEDIUM II

This medium was extracted automatically and has not been curated by experts, yet.

Culture medium recipe

Vol.
×
Medium for Chlorobium limicola DSM 245

* Modification: Omit Yeast extract, Dextrose, and Pyruvic acid sodium salt.

Growth conditions: 7-14 days 25 °C

Main sol. 29 #

ml
Solution occurs in 1 other media (first defined in Medium 29).
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 B 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/l - -
Vitamin B12 solution 1 ml/l - -
6 Adjust pH to 6.8-7.1 with 1M Na2CO3 (filter-sterilized).
7 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).

Neutralized sulfide solution 3% (w/v) #

100
Solution occurs in 20 other media (first defined in Medium 28).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 3 g - -
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.

Trace element solution SL-10 B #

1000
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 - -
FeSO4 x 7 H2O 1.5 g - -
ZnCl2 70.0 mg - -
MnCl2 x 4 H2O 100.0 mg - -
H3BO3 300.0 mg - -
CoCl2 x 6 H2O 190.0 mg - -
CuCl2 x 2 H2O 2.0 mg - -
NiCl2 x 6 H2O 24.0 mg - -
Na2MoO4 x 2 H2O 36.0 mg - -

Solution A #

460
Solution occurs in 6 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 - -
Yeast extract* 0.25 g - -
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.

Sulfide solution, 1.5% #

135
Solution occurs in 6 other media (first defined in Medium 28).
Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 2 g - -
Distilled water 135 ml - -
1 Prepare in a screw-cap bottle, bubble with N2 to replace air, close tightly and autoclave.

Bicarbonate solution #

50
Solution occurs in 6 other media (first defined in Medium 28).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaHCO3 1.5 g - -
H2O 50.0 ml - -
1 Bubble with CO2 and filter sterilize into sterile, gas-tight, 100 ml screw-cap bottle.

Resazurin solution #

450
Solution occurs in 6 other media (first defined in Medium 28).
Equipment needed: Ice bath Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Resazurin
(0,1%) 		0.5 ml - -
Distilled water 450.0 ml - -
1 Cool to room temperature under an atmosphere of N2/CO2 in an ice bath.
2 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.

Pfennig's heterotrophic salts solution B #

26
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ammonium chloride 0.35 g - -
Ammonium acetate 0.25 g - -
Pyruvic acid sodium salt* 0.25 g - -
Dextrose* 0.25 g - -
MgSO4 x 7 H2O 0.50 g - -
KCl 0.35 g - -
KH2PO4 0.35 g - -
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.

Vitamin B12 solution #

100
Solution occurs in 7 other media (first defined in Medium 28).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin B12 0.01 g - -
Distilled water 100.00 ml - -
1 Filter sterilized

Trace element solution SL-10 B #

1000
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(25%) 		7.7 ml - -
FeCl2 x 4 H2O 1.5 g - -
ZnCl2 70.0 mg - -
MnCl2 x 4 H2O 100.0 mg - -
H3BO3 300.0 mg - -
CoCl2 x 6 H2O 190.0 mg - -
CuCl2 x 2 H2O 2.0 mg - -
NiCl2 x 6 H2O 24.0 mg - -
Na2MoO4 x 2 H2O 36.0 mg - -
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.

Metadata

Last modified: 30.06.26
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Defined medium
Final pH: 6.8 - 7.1
Equipment needed: Autoclave, Filter, Ice bath
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Associated strains

All strains for this medium: 12
Strains with modifications: 3
Chlorobium limicola DSM 245

  Yeast extract
  Dextrose
  Pyruvic acid sodium salt

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 12 strains BacDive

phyla 2 12 strains
nutrition phototroph 3 strains
oxygen anaerobe 11 strains
oxygen obligate anaerobe 3 strains
temperature mesophilic 10 strains
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