1503
1503 .
CULTIVATION MEDIUM FOR CHLAMYDOPHILA PECORUM
This medium was extracted automatically and has not been curated by experts, yet.
Culture medium recipe
Vol.
×
Medium for Chlamydia pecorum DSM 27092
* Modification: Plus 5% CO2
The modifications for this strain were extracted automatically and have not been curated by experts, yet.
Growth conditions: 37 °C
Main sol. 1503 #
ml
Equipment needed: Filter
Centrifuge
| 1 Cultivation of CACO-2 cells (ACC 169) | |||||
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| MEM-Medium | 45.0 | ml | - | - | |
| Fetal bovine serum | 5.0 | ml | - | - | |
|
Aminoacids
(100 x) |
0.5 | ml | - | - | |
| 2 Filter-sterilize (0.2 µm) and keep no longer than 4 weeks. Store at room temperature to facilitate detection of contamination. | |||||
| 3 Prepare a 25 cm2 flask and seed cells according to standard protocols (see DSMZ catalogue for ACC 169). Incubate at 37°C plus 5% CO2. When a confluent layer has formed, infection can be carried out. | |||||
| 4 Exchange medium with 6 ml of Infection Medium (as above with the addition of 1 µg/ml cycloheximide (final concentration)) and add 500 - 1000 µl of EB stock solution (thawed quickly to 37°C). | |||||
| 5 Centrifuge for 1 h onto the cell layer at 1600 rpm at 20°C. | |||||
| 6 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. | |||||
Metadata
| Last modified: | 24.02.22 |
| Source: | DSMZ |
| Taxonomic range: | Bacteria |
| Medium type: | Complex medium |
| Final pH: | n.d. |
| Equipment needed: | Centrifuge, Filter |
| Edit in Medium builder | |
Gas composition
- 5 % CO2
Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .
Molecular composition
| Compound | Conc. [g/L] | Conc. [mM] |
|---|---|---|
| Aminoacids | - | - |
| Fetal bovine serum | - | - |
| MEM-Medium | - | - |
