Chlorogonium elongatum Dangeard

1. To thaw a frozen ampule, place in a 35°C water bath, until thawed (2-3 min). Immerse the ampule just sufficiently to cover the frozen material.  Do not agitate the ampule.
2. Immediately after thawing, aseptically transfer the entire contents to a single 16 x 125 mm screw-capped test tube containing 5 mL of ATCC Medium 5 broth (or alternatively, 5 mL ATCC Medium 351 supplemented with 0.1% Na Acetate).  Allow the tube to stand upright for one to several hours (or centrifuge very lightly for 3-5 min) to allow most cells to settle to the tube base.
3. Gently remove most of the supernatant (save in a secondary tube), then resuspend the remaining cells in additional fresh medium to a total volume of 5-6 mL.
4. Incubate tubes on a 15° horizontal slant with the cap screwed on loosely (loosened one half turn) at 20-25°C under a 14 hour light (~50 µEinsteins/m²/s irradiance)/10 hour dark cycle.
   Note: Some algae may grow poorly or not at all when recovered from the frozen state in broth media.  In such cases, the algae should initially be recovered on agar media before eventually transferring to broth culture.

1. Inoculate a tube of fresh broth medium with 0.25 mL from a growing culture at or near peak density.
2. Incubate tubes on a 15° horizontal slant with the cap screwed on loosely (loosened one half turn) at 20-25°C under a 14 hour light (~50 °Einsteins/m²/s irradiance)/10 hour dark cycle.
3. Subculture as necessary (i.e., typically every 14-21 days in broth media).

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