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DSMZ

DSMZ 1761 . PFENNIG'S MEDIUM III

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

Culture medium recipe

Vol.
×

Main sol. 1761 #

ml
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 46.0 ml - -
Solution B 2.6 ml - -
Solution C 5.0 ml - -
Solution D 45.0 ml - -
Solution E 2.6 ml - -
Solution F 0.1 ml - -
Solution G 1.0 ml - -
1 Adjust pH to 6.0.
2 Mix solution D, C and E in the cotton-stoppered Erlenmeyer flask. Bubble with CO2 in an ice bath under sterile conditions (5-10 min).
3 Fill 50 ml of the mixed solution (C,D,E) into each 100 ml bottle of solution A (for a total volume of approx. 96 ml).
4 Before using, add 2,6 ml solution B (Na2S x 9 H2O), 0.1 ml solution F (Vitamin B12) and 1 ml of solution G (2,5% Ammonium-acetate and Magnesium-acetate) to each 100ml bottle.
5 Adjust the pH with filter-sterilised 1M Na2CO3 to 7.1-7.3.
6 Fill in sterile, N2 gassed screw-cap tubes under N2 gas.
7 For the growth of DSM 11081 feed the culture with neutralized sulfide feeding solution regularly (approx. every 3 days). 0,5-1 ml/ 100 mL

Solution A #

460
Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
CaCl2 x 2 H2O 0.25 g 0.543 3.697
Distilled water 460.00 ml - -
1 Fill 10x 46ml in 100 ml screw-cap bottles. Bubble with N2/CO2 and autoclave at 121°C for 15 min.

Solution B #

135
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 C #

50
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 D #

450
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. Cool to room temperature under an atmosphere of N2/CO2 in an ice bath (10-15 min).

Solution E #

26
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ammonium chloride 0.40 g 15.385 287.617
MgSO4 x 7 H2O 0.50 g 19.231 78.023
KCl 0.34 g 13.077 175.411
KH2PO4 0.34 g 13.077 96.094
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 F #

100
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin B12 2 mg 0.02 0.015
Distilled water 100 ml - -
1 Filter sterilized

Solution G #

100
Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ammonium acetate 2.5 g 25 324.338
Magnesium acetate 2.5 g 25 175.57
Distilled water 100.0 ml - -
1 Autoclave at 121°C for 15 min.

Trace element solution SL-12 B #

1000
Equipment needed: Ice bath Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Distilled water 1000.0 ml - -
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

Neutralized sulfide solution 3% (w/v) #

100
Solution occurs in 18 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: 15.04.22
Source: DSMZ
Medium type: Defined medium
Final pH: 6.0
Equipment needed: Autoclave, Filter, Ice bath
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Associated strains

All strains for this medium: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaHCO3 1.5 17.856
MgSO4 x 7 H2O 0.5 2.029
Ammonium chloride 0.4 7.478
Na2S x 9 H2O 0.385 1.604
KCl 0.34 4.561
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature thermophilic 1 strain