chlamydia_serpentis_poikilotherma_medium

Overview

External ontology

mediadive.medium:1793 (source link unavailable) — Chlamydia serpentis & poikilotherma Medium

Applications

• Microbial cultivation

Notes

Source: DSMZ

Composition

flowchart LR
    CultureMech_001238["chlamydia_serpentis_poikilotherma_medium"]
    ing0_MEM_Medium["MEM-Medium"]
    CultureMech_001238 --> ing0_MEM_Medium
    ing1_Fetal_bovine_serum["Fetal bovine serum"]
    CultureMech_001238 --> ing1_Fetal_bovine_serum
    ing2_D_Glucose["D-Glucose"]
    CultureMech_001238 --> ing2_D_Glucose
    CHEBI_17634["CHEBI:17634"]
    ing2_D_Glucose -.-> CHEBI_17634
    ing3_Demineralized_water["Demineralized water"]
    CultureMech_001238 --> ing3_Demineralized_water
    CHEBI_15377["CHEBI:15377"]
    ing3_Demineralized_water -.-> CHEBI_15377
    ing4_Cycloheximid["Cycloheximid"]
    CultureMech_001238 --> ing4_Cycloheximid
    CHEBI_27641["CHEBI:27641"]
    ing4_Cycloheximid -.-> CHEBI_27641

Ingredients (5)

Preparation

1. Filter-sterilize (0.2 µm) and keep no longer than 4 weeks. Store at room temperature to facilitate detection of contamination.
2. Prepare a 25 cm2 flask and seed cells according to standard protocols. Incubate at 37°C plus 5% CO2. When a confluent layer has formed, infection can be carried out.
3. Exchange medium with Infection Medium (as above with the addition of cycloheximide) and add 500 - 1000 µl of EB stock solution (thawed quickly to 37°C).
4. Centrifuge for 1 h onto the cell layer at 1000 x g at >20°C.
5. Incubate at 37°C + 5% CO2. Control cells daily and look for inclusions. Not all Chlamydiae form well-visible inclusions, ultimately, immunofluorescence or in situ-hybridization techniques are necessary to visualize inclusions.
6. Filtrate.
7. Filtrate.

Target organisms

No target organisms are recorded.

Provenance