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1. Harvest cells from a culture which is at or near peak density by centrifugation at 1,300 g for 5 min.
2. Adjust concentration of cells to 2 x 10⁷/mL in fresh medium.
3. While cells are centrifuging prepare a 10% (v/v) solution of sterile DMSO in fresh medium (broth).  The DMSO solution when first prepared will warm up due to chemical heat. The solution should be allowed to return to room temperature prior to use.
4. Mix the cell preparation and the DMSO solution in equal portions. The final concentration will be 10⁷ cells/mL and 5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should be no more than 15 min.
5. Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
6. Place the ampules in a Nalgene 1°C freezing apparatus.  Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.  (The cooling rate in this apparatus is approximately -1°C/min.)  
7. Store in either the vapor or liquid phase of a nitrogen refrigerator.
8. To thaw a frozen ampule, place it in a 35°C water bath such that the lip of the ampule remains above the water line. Thawing time is approximately 2 to 3 minutes.  Do not agitate the ampule.  Do not leave ampule in water bath after thawed.
9. Immediately after thawing, do not leave in the water bath, aseptically transfer the contents of the ampule into a fresh tube of ATCC medium 1011. 
10. Incubate vertically at 25°C with the cap screwed on tightly.
11. Maintain as described above. 

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Galinari S, Camargo EP. Trypanosomatid protozoa: survey of acetylornithinase and ornithine acetyltransferase. Exp. Parasitol. 46: 277-282, 1978. PubMed: 569594

Fish WR, et al. The cyclopropane fatty acid of trypanosomatids. Mol. Biochem. Parasitol. 3: 103-115, 1981. PubMed: 7254247

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Fish WR, et al. Some Phytomonas and Herpetomonas species form unique iso-branched polyunsaturated fatty acids. Mol. Biochem. Parasitol. 5: 1-18, 1982. PubMed: 7062937

Alfieri SC, Camargo EP. Trypanosomatidae: isoleucine requirement and threonine deaminase in species with and without endosymbionts. Exp. Parasitol. 53: 371-380, 1982. PubMed: 6806116

Camargo EP, et al. Electrophoretic analysis of endonuclease-generated fragments of k-DNA, of esterase isoenzymes, and of surface proteins as aids for species identification of insect trypanosomatids. J. Protozool. 29: 251-258, 1982. PubMed: 6284925

type strain

Nucleotide (GenBank) : U01007 Herpetomonas muscarum muscarum kinetoplast 9S rRNA gene.

Nucleotide (GenBank) : U01011 Herpetomonas muscarum muscarum kinetoplast 12S rRNA gene.

Nucleotide (GenBank) : U01015 Herpetomonas muscarum muscarum 16S-like small subunit rRNA.