Mycoplasma Testing

Mycoplasma testing using MycoAlert® System

The Tissue Culture Facility performs mycoplasma testing for your lab’s cultured cells. The TCF offers one day turnaround on results, and depending on work load, within an hour.

The new MycoAlert® System detects mycoplasma in cell cultures using the sensitive luminescent assay of ATP. The assay is a selective biochemical test that exploits the activity of mycoplasmal enzymes. The viable mycoplasma are lysed and the enzymes react with the MycoAlert® substrate, catalyzing the conversion of ADP to ATP. By measuring ATP before and after addition of the substrate, a ratio is obtained which indicates the presence or absence of mycoplasma.

This assay can be performed at every passage to ensure that the cultures remain uncontaminated.

Preparation of Samples for Mycoplasma Testing

  1. 1. Samples of the culture medium should be taken before any further processing steps, e.g. trypsinization.
  2. 2. Cultures should be free of any antibiotics or antimycotics for 2-3 passages, and approximately 80% confluent.
  3. 3. Transfer 10 ml of cell culture or culture supernatant into a 15 ml centrifuge tube and pellet any cells at 1500 rpm (200 x g) for 5 minutes.
  4. 4. Transfer 10 ml of cleared supernatant into a sterile tube and label it.
  5. 5. Bring samples to the Tissue Culture Facility and fill out a request form.