1. Thaw and mix Mycoplasma Elimination Reagent (Myco-1+2) and penicillin-free and streptomycin-free culture medium thoroughly.
2. Dilute each reagent (Myco-1+2) to 1/100 respectively.

1. When performing cell passage or continuing to culture with mycoplasma contaminated cells, replace the culture medium with Myco-1-added medium and maintain the contaminated cells in the medium for 4 days.
2. After 4 days, replace the medium with Myco-2-added medium and maintain the contaminated cells in the medium for 3 days. For most mycoplasma contamination, it may be necessary to repeat this treatment cycle 1-2 times until the mycoplasma is removed.
3. 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, repeat the treatment cycle another 1-2 times.

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 a clean bench.
• It is recommended to add this product to the 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.
• For mycoplasma treatment, Myco-1 and Myco-2 should be used sequentially for best results.
• Mycoplasma Elimination Reagent (Myco-1+2) has been tested in a variety of cell lines, including HepG2, HEK-293T, HEK-293, HeLa, N2a, A549, MCF-7, CHO, K562, NIH/3T3, Sp2/0, BHK-21, VERO, U937, and SY5Y.
• This product can be used on primary cells with low cytotoxicity. If the cells are very sensitive, it is recommended to reduce the concentration.
• Once the cells have been contaminated and treated, it is very easy for re-contamination to occur. The use of a Mycoplasma prevention reagent is recommended (abx098886) to prevent any further contamination.