Induction of Conjugation in Tetrahymena Protocol Mature Tetrahymena cells of opposite mating types are starved under appropriate salt conditions. The mating types are then combined to costimulate through cell-cell interaction. Loose pairs and then firm, irreversible pairs of cells of opposite mating types form. This method consistently results in a high percentage of pairing (usually greater than 80%) and good synchrony.
Isolation and Purification of Tetrahymena Nuclei Protocol Protocol describes how to allow the isolation of nuclei from all stages of the Tetrahymena life cycle in high yield with a high degree of purity. This method gives highly purified populations of both micronuclei and macronuclei.
Short-Term Storage of Tetrahymena Cultures Protocol A number of methods can be used for storage of unfrozen Tetrahymena cultures in the laboratory. Cells that are maintained using the short-term storage are described in this protocol.
Transformation of Tetrahymena thermophila by Electroporation Protocol Protocol describes a method for transformation of the Tetrahymena using electroporation. The vector is electroporated into cells after mating, where it is incorporated into the DNA of developing macronuclei. Because T. thermophila can be propagated indefinitely without conjugation, transformation of the macronucleus provides a way to obtain stable somatic transformants. DNA vectors transformed using this protocol include those containing drug-resistant versions of Tetrahymena genes.
