Mycoplasmoides genitalium (Tully et al.) Gupta et al.

1. We recommend that you use T-flasks in the propagation of Mycoplasma genitalium so that a larger surface area will be available for the cells to adhere to the glass or plastic surface. Approximately 10 mL of Medium #988 is placed in 25 cm² flasks, and 200 mL can be placed in 125 cm² flasks if larger amounts are desired.
2. Add an additional 5% heat-inactivated fetal bovine serum to medium #988. Open the vial according to enclosed instructions.
3. Transfer the contents of the vial to a 25 cm² flask.
4. Incubate the flask in a horizontal position at 37°C for 5-7 days. This strain can be slow to establish growth, however subsequent transfers should grow faster once primary growth is established. Observe the flask for flocs in the medium and for cells adhering to the bottom of the flask. Growth is also indicated by an indicator change from red to yellow.
5. When the medium has turned yellow, loosen adherent cells with a suitable, sterile scraper. Use a 10% inoculum when transferring this item to ensure healthy growth in sub-cultures.

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