I want to check presence of mycoplasma in cell culture by broth/agar method.
Dear Surendra Gohil
i'm not able to answer to your specific question but i can suggest to you a simple PCR protocoll (in the following link) that allow you to check for presence of micoplasma on any cell culture media that you would like to check for it.
https://www.blogger.com/video.g?token=AD6v5dw7NqhRBr-g9xdXI151JBzQwSayjswhAYtYQ-Q_5Q6WPgZBgm_Mkqnq8LRgwlQ_5WZoyIlI6xYbi6Ha4XFXgxurdyOnJ5a3C7MicdqjrKBBrUX3sPrOUpcAaxm0VlTISU_AiQ
good luck
Manuele
Hello Surendra Gohil
The microbiological culture method is a gold standard to detect every kind of mycoplasma contamination in cell culture without considering the origin and species of mycoplasma.
In this method, the supernatant from the cell culture is added to the liquid or semi-solid medium (containing nutrients essential for mycoplasma growth). Infected cell culture will show growth both in the liquid broth and on the agar plates. A sample of cell culture supernatant is cultivated on a mycoplasma agar medium and incubated in an aerobic condition at 37ºC for about two weeks. Positive samples on agar plates will show small colonies similar to fried eggs with a diameter of 100-400 µm.
Preparation and components of the media to grow mycoplasma are described in the references given below.
https://pubmed.ncbi.nlm.nih.gov/20431538/
Article Sensitivity and specificity of five different Mycoplasma det...
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