pfennigs_medium_ii_with_salt

Overview

External ontology

mediadive.medium:40 (source link unavailable) — PFENNIG'S MEDIUM II WITH SALT

Applications

• Microbial cultivation

High-metal flag

True

Notes

Source: DSMZ | Link: https://www.dsmz.de/microorganisms/medium/pdf/DSMZ_Medium40.pdf

Composition

flowchart LR
    CultureMech_001518["pfennigs_medium_ii_with_salt"]
    ing0_NaCl["NaCl"]
    CultureMech_001518 --> ing0_NaCl
    CHEBI_26710["CHEBI:26710"]
    ing0_NaCl -.-> CHEBI_26710
    ing1_CaCl2_x_2_H2O["CaCl2 x 2 H2O"]
    CultureMech_001518 --> ing1_CaCl2_x_2_H2O
    CHEBI_86158["CHEBI:86158"]
    ing1_CaCl2_x_2_H2O -.-> CHEBI_86158
    ing2_Yeast_extract["Yeast extract"]
    CultureMech_001518 --> ing2_Yeast_extract
    ing3_NaHCO3["NaHCO3"]
    CultureMech_001518 --> ing3_NaHCO3
    CHEBI_32139["CHEBI:32139"]
    ing3_NaHCO3 -.-> CHEBI_32139
    ing4_Resazurin["Resazurin"]
    CultureMech_001518 --> ing4_Resazurin
    CHEBI_8806["CHEBI:8806"]
    ing4_Resazurin -.-> CHEBI_8806
    ing5_Ammonium_chloride["Ammonium chloride"]
    CultureMech_001518 --> ing5_Ammonium_chloride
    CHEBI_31206["CHEBI:31206"]
    ing5_Ammonium_chloride -.-> CHEBI_31206
    ing6_Ammonium_acetate["Ammonium acetate"]
    CultureMech_001518 --> ing6_Ammonium_acetate
    CHEBI_62947["CHEBI:62947"]
    ing6_Ammonium_acetate -.-> CHEBI_62947
    ing7_Pyruvic_acid_sodium_salt["Pyruvic acid sodium salt"]
    CultureMech_001518 --> ing7_Pyruvic_acid_sodium_salt
    CHEBI_50144["CHEBI:50144"]
    ing7_Pyruvic_acid_sodium_salt -.-> CHEBI_50144
    ing8_Dextrose["Dextrose"]
    CultureMech_001518 --> ing8_Dextrose
    CHEBI_17634["CHEBI:17634"]
    ing8_Dextrose -.-> CHEBI_17634
    ing9_MgSO4_x_7_H2O["MgSO4 x 7 H2O"]
    CultureMech_001518 --> ing9_MgSO4_x_7_H2O
    CHEBI_31795["CHEBI:31795"]
    ing9_MgSO4_x_7_H2O -.-> CHEBI_31795
    ing10_KCl["KCl"]
    CultureMech_001518 --> ing10_KCl
    CHEBI_32588["CHEBI:32588"]
    ing10_KCl -.-> CHEBI_32588
    ing11_KH2PO4["KH2PO4"]
    CultureMech_001518 --> ing11_KH2PO4
    CHEBI_63036["CHEBI:63036"]
    ing11_KH2PO4 -.-> CHEBI_63036
    ing12_Vitamin_B12["Vitamin B12"]
    CultureMech_001518 --> ing12_Vitamin_B12
    CHEBI_176843["CHEBI:176843"]
    ing12_Vitamin_B12 -.-> CHEBI_176843
    ing13_Na2S_x_9_H2O["Na2S x 9 H2O"]
    CultureMech_001518 --> ing13_Na2S_x_9_H2O
    CHEBI_76209["CHEBI:76209"]
    ing13_Na2S_x_9_H2O -.-> CHEBI_76209
    ing14_Na2_EDTA["Na2-EDTA"]
    CultureMech_001518 --> ing14_Na2_EDTA
    CHEBI_64734["CHEBI:64734"]
    ing14_Na2_EDTA -.-> CHEBI_64734
    ing15_FeSO4_x_7_H2O["FeSO4 x 7 H2O"]
    CultureMech_001518 --> ing15_FeSO4_x_7_H2O
    CHEBI_75836["CHEBI:75836"]
    ing15_FeSO4_x_7_H2O -.-> CHEBI_75836
    ing16_CoCl2_x_6_H2O["CoCl2 x 6 H2O"]
    CultureMech_001518 --> ing16_CoCl2_x_6_H2O
    CHEBI_53503["CHEBI:53503"]
    ing16_CoCl2_x_6_H2O -.-> CHEBI_53503
    ing17_MnCl2_x_2_H2O["MnCl2 x 2 H2O"]
    CultureMech_001518 --> ing17_MnCl2_x_2_H2O
    CHEBI_131395["CHEBI:131395"]
    ing17_MnCl2_x_2_H2O -.-> CHEBI_131395
    ing18_ZnCl2["ZnCl2"]
    CultureMech_001518 --> ing18_ZnCl2
    CHEBI_49976["CHEBI:49976"]
    ing18_ZnCl2 -.-> CHEBI_49976
    ing19_NiCl2_x_6_H2O["NiCl2 x 6 H2O"]
    CultureMech_001518 --> ing19_NiCl2_x_6_H2O
    CHEBI_34887["CHEBI:34887"]
    ing19_NiCl2_x_6_H2O -.-> CHEBI_34887
    ing20_Na2MoO4_x_2_H2O["Na2MoO4 x 2 H2O"]
    CultureMech_001518 --> ing20_Na2MoO4_x_2_H2O
    CHEBI_75213["CHEBI:75213"]
    ing20_Na2MoO4_x_2_H2O -.-> CHEBI_75213
    ing21_H3BO3["H3BO3"]
    CultureMech_001518 --> ing21_H3BO3
    CHEBI_33118["CHEBI:33118"]
    ing21_H3BO3 -.-> CHEBI_33118
    ing22_CuCl2_x_2_H2O["CuCl2 x 2 H2O"]
    CultureMech_001518 --> ing22_CuCl2_x_2_H2O
    CHEBI_86318["CHEBI:86318"]
    ing22_CuCl2_x_2_H2O -.-> CHEBI_86318

Ingredients (23)

Preparation

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.
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.
6. 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).
7. Adjust pH to 6.8-7.1 with 1M Na2CO3 (filter-sterilized).
8. Fill 10x 46ml in 100 ml screw-cap bottles. Bubble with N2/CO2 and autoclave 121°C 15 min.
9. Prepare in a screw-cap bottle, bubble with N2 to replace air, close tightly and autoclave.
10. Bubble with CO2 and filter sterilize into sterile, gas-tight, 100 ml screw-cap bottle.
11. 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.
12. Cool to room temperature under an atmosphere of N2/CO2 in an ice bath.
13. Filter sterilize into sterile, gas-tight, 100 ml screw-cap bottle.
14. Filter sterilized
15. 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.
16. Adjust the pH with filter-sterilised 1M Na2CO3 to 7.1-7.3.
17. If needed, aliquot into sterile, N₂ gassed screw-cap tubes under N₂ gas.
18. 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.
19. 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).
20. Prepare the following solutions (resazurin, bicarbonate and Pfennig's heterotrophic salts) and sterilize as given below.
21. 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.
22. Fill 50 ml of this mixture to each bottle of solution A (46 mL + 50 mL).
23. Before use, add 4 ml sulfide solution (1.5%) and 0.1 ml Vitamin B12 solution to each 100 mL bottle.
24. 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).
25. Prepare in a screw-cap bottle, bubble with N2 to replace air, close tightly and autoclave.
26. Bubble with CO2 and filter sterilize into sterile, gas-tight, 100 ml screw-cap bottle.
27. 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.
28. Cool to room temperature under an atmosphere of N2/CO2 in an ice bath.
29. Filter sterilize into sterile, gas-tight, 100 ml screw-cap bottle.
30. Filter sterilized
31. 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.
32. Adjust pH to 6.0.

Target organisms

No target organisms are recorded.

Provenance