semi_solid_heterotrophic_growth_medium_for_alphaproteobacteria

Overview

External ontology

jcm.grmd:1430 (source link unavailable) — SEMI-SOLID HETEROTROPHIC GROWTH MEDIUM FOR ALPHAPROTEOBACTERIA

Applications

• Microbial cultivation

pH

6.7

Notes

Source: JCM | Link: https://www.jcm.riken.jp/cgi-bin/jcm/jcm_grmd?GRMD=1430

Composition

flowchart LR
    CultureMech_015865["semi_solid_heterotrophic_growth_medium_for_alphaproteobacteria"]
    ing0_NaCl["NaCl"]
    CultureMech_015865 --> ing0_NaCl
    CHEBI_26710["CHEBI:26710"]
    ing0_NaCl -.-> CHEBI_26710
    ing1_MgCl2_6H2O["MgCl2·6H2O"]
    CultureMech_015865 --> ing1_MgCl2_6H2O
    CHEBI_86345["CHEBI:86345"]
    ing1_MgCl2_6H2O -.-> CHEBI_86345
    ing2_Na2SO4["Na2SO4"]
    CultureMech_015865 --> ing2_Na2SO4
    CHEBI_32149["CHEBI:32149"]
    ing2_Na2SO4 -.-> CHEBI_32149
    ing3_KCl["KCl"]
    CultureMech_015865 --> ing3_KCl
    CHEBI_32588["CHEBI:32588"]
    ing3_KCl -.-> CHEBI_32588
    ing4_CaCl2_2H2O["CaCl2·2H2O"]
    CultureMech_015865 --> ing4_CaCl2_2H2O
    CHEBI_86158["CHEBI:86158"]
    ing4_CaCl2_2H2O -.-> CHEBI_86158
    ing5_NH4Cl["NH4Cl"]
    CultureMech_015865 --> ing5_NH4Cl
    CHEBI_31206["CHEBI:31206"]
    ing5_NH4Cl -.-> CHEBI_31206
    ing6_NaHCO3["NaHCO3"]
    CultureMech_015865 --> ing6_NaHCO3
    CHEBI_32139["CHEBI:32139"]
    ing6_NaHCO3 -.-> CHEBI_32139
    ing7_HEPES["HEPES"]
    CultureMech_015865 --> ing7_HEPES
    CHEBI_46756["CHEBI:46756"]
    ing7_HEPES -.-> CHEBI_46756
    ing8_Yeast_extract__BD_Difco_["Yeast extract (BD-Difco)"]
    CultureMech_015865 --> ing8_Yeast_extract__BD_Difco_
    ing9_Disodium_succinate["Disodium succinate"]
    CultureMech_015865 --> ing9_Disodium_succinate
    CHEBI_63675["CHEBI:63675"]
    ing9_Disodium_succinate -.-> CHEBI_63675
    ing10_Modified_Wolfe_s_mineral_solution__see_Medium_No__915__["Modified Wolfe’s mineral solution (see Medium No. 915 )"]
    CultureMech_015865 --> ing10_Modified_Wolfe_s_mineral_solution__see_Medium_No__915__
    ing11_Resazurin["Resazurin"]
    CultureMech_015865 --> ing11_Resazurin
    CHEBI_8806["CHEBI:8806"]
    ing11_Resazurin -.-> CHEBI_8806
    ing12_Distilled_waterv["Distilled waterv"]
    CultureMech_015865 --> ing12_Distilled_waterv
    ing13_Agarose["Agarose"]
    CultureMech_015865 --> ing13_Agarose
    CHEBI_2511["CHEBI:2511"]
    ing13_Agarose -.-> CHEBI_2511
    ing14_0_5_M_Potassium_phosphate_buffer__pH_7_0_["0.5 M Potassium phosphate buffer (pH 7.0)"]
    CultureMech_015865 --> ing14_0_5_M_Potassium_phosphate_buffer__pH_7_0_
    ing15_Vitamin_solution___see_Medium_No__915__["Vitamin solution* (see Medium No. 915 )"]
    CultureMech_015865 --> ing15_Vitamin_solution___see_Medium_No__915__
    ing16_0_01_M_FeSO4_7H2O_solution__in_0_02_N_HCl_["0.01 M FeSO4·7H2O solution (in 0.02 N HCl)"]
    CultureMech_015865 --> ing16_0_01_M_FeSO4_7H2O_solution__in_0_02_N_HCl_
    ing17_5__L_Cysteine_HCl_H2O_solution["5% L-Cysteine·HCl·H2O solution"]
    CultureMech_015865 --> ing17_5__L_Cysteine_HCl_H2O_solution

Ingredients (18)

Preparation

1. Mix components thoroughly, except agarose, and adjust pH to 6.7.
2. Add agarose to the medium and autoclave under a N2 gas atmosphere.
3. After cooling to 50C, add the following solutions to the medium aseptically and anaerobically (autoclaved or *filter-sterilized).
4. Keep the medium temperature at 50 C until the medium is distributed into sterile screw-capped test tubes as described below: Adjust pH to 7.0 with sterile HCl or NaOH solution.
5. Dispense the medium to sterile screw-capped test tubes (typically 11 ml medium into sterile Hungate tubes) under air and cap.
6. Allow to stand until the medium is solidified and a O2-gradient formed (as evidenced by a pink upper oxic zone and a colorless lower anoxic zone).
7. Inoculate cells just below the pink-colorless interface in the medium tube using a syringe.
8. Comment: The medium tubes can be stocked under a N2 gas atmosphere.
9. In this case, prior to use, fill the head space with air and allow to stand until the O2 gradient is formed.

Target organisms

No target organisms are recorded.

Provenance