Procedure:

1. Preparation of the culture medium: Thaw and mix penicillin-free and streptomycin-free culture medium thoroughly. To 99 ml of this culture medium, add 1 ml of Mycoplasma Elimination Reagent (Myco-3).
2. When performing cell passage or culturing with mycoplasma contaminated cells, replace the culture medium with Myco-3-added medium and maintain the contaminated cells in the medium for 3 days.
3. Continue the treatment for a total of 10-14 days by changing with the medium (containing Myco-3) every 2-3 days. For most mycoplasma contamination, it may be necessary to maintain the cells in the medium for 10-14 days until the mycoplasma is removed.
4. During the process, the cells can be tested for mycoplasma contamination to detect the removal efficiency. In general, mycoplasma will be removed after 2 weeks of treatment. If the culture still tests positive, continue the treatment for another 3-7 days.

Notes:

• Prior to use, ensure that the cap of the bottle is sealed. Thaw to room temperature before use. Mix gently and use 70% ethanol to wipe the bottle surface, and then place it on the clean bench.
• Add this product to fresh culture medium.
• For the fast proliferation of cell lines, in order to reduce the number of passage, it is recommended to reduce the density of cells or reduce the concentration of serum in medium.
• If the cells are sensitive to Myco-3, it is recommended to reduce the concentration of the reagent.
• It is recommended to use a mycoplasma prevention reagent, such as abx098886 once treated to prevent future contamination.