Herpetomonas muscarum muscarum Rogers and Wallace

1. To thaw a frozen ampule, place in a 35°C water bath, until thawed (2-3 min). Immerse the ampule just sufficient to cover the frozen material.  Do not agitate the ampule.
2. Immediately after thawing, aseptically transfer contents to a screw-capped borosilicate test tube containing ATCC Medium 1011. Incubate the tube vertically at 25°C with the cap screwed on tightly.

1. When the culture has reached or is near peak density, invert tube 10 times and aseptically transfer a drop from a Pasteur pipette (0.05 mL) to another test tube containing fresh ATCC medium 1011.
2. Incubate the culture vertically at 25°C with the cap screwed on tightly. 
3. Transfer the culture every 3-4 days as described in step 1.  The transfer interval will depend on the quantity of the inoculum and the quality of the medium.  This should be empirically determined by examining the culture on a daily basis until the growth cycle has stabilized.

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