Laboratory Model Organisms Protocols

Culturing Marine Euplotids Using Dunaliella as a Food Source Protocol Protocol describes the culture of marine euplotids using Dunaliella salina or D. tetiolecta as a food organism. Dunaliella tolerate a wide range of salinity, thus they are fairly easy to grow in the lab using artificial sea salts.

Growth and Concentration of Chlorogonium for Culturing Freshwater Hypotrichs Protocol Protocol describes the growth and concentration of the alga Chlorogonium elongatum as a food source for culturing freshwater hypotrichs. Most freshwater hypotrichs (including Oxytricha nova, O. fallax, and O. trifallax; Euplotes aediculatus and E. eurystomous; and Stylonychia lemnae) can be grown to high density with Chlorogonium as the food organism. A similar regimen can be used to prepare other food sources such as Tetrahymena or bacteria (e.g., Aerobacter aerogenes).

Immunohistochemistry on Cryosections from Embryonic and Adult Zebrafish Eyes Protocol Protocol describes methods for the fixation, cryosectioning, and immunohistochemical detection of proteins in embryonic and adult zebrafish eyes. The methods described can easily be adapted for use on other zebrafish tissues.

Maintenance of Stocks of Marine Hypotrichs Protocol Protocol describes the preparation of stocks of marine hypotrichs for long-term storage. Marine euplotids are maintained by serial passage of tube stocks.

Mass Culture of Freshwater Hypotrichs Protocol Protocol describes the culture of freshwater hypotrichs using the alga Chlorogonium elongatum as a food source.

Pristionchus pacificus Genetic Protocols Genetic Protocols for Pristionchus pacificus. Includes: Freezing worms; EMS mutagenesis; Psoralen mutagenesis; Construction of deletion libraries to generate P. pacificus gene knock-outs; Designing primers for the gene of interest; RNAi and morpholino by injection.

Refeeding Marine Euplotids with Bacteria Protocol Feeding euplotids with algae can lead to asynchronous cell starvation and vastly different cell sizes within a culture. Asynchronous starvation also leads to different levels of mating competence. Furthermore, algal pigment remnants can interfere with many applications (e.g., fluorescence microscopy).