Dictyostelium discoideum Raper

1. Open an ampoule according to enclosed instructions.
2. From a single test tube of sterile distilled water (5 to 6 mL), withdraw approximately 0.5 to 1.0 mL with a sterile pipette and apply directly to the pellet. Stir to form a suspension.
3. Aseptically transfer the suspension back into the test tube of sterile distilled water.
4. Let the test tube sit at room temperature (25°C) undisturbed for at least 2 hours; longer (e.g., overnight) rehydration might increase viability of some fungi.
5. Mix the suspension well. Use 1.0 mL to inoculate recommended freshly-made and wetted solid medium and freshly-made liquid medium.  Agar plates should be overlaid with 2 mL of a liquid medium, either #671 or #1034 (with 10% fetal bovine serum included). Include a control that receives no inoculum.
6. Incubate the inoculum at the propagation conditions recommended.
7. Inspect for growth of the inoculum/strain regularly. The sign of viability is noticeable typically after 2-3 days of incubation. However, the time necessary for significant growth will vary. 
8. Subculture from stationary liquid cultures after one week.

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