Methods

These are worm methods optimized for Pristionchus pacificus. Common C. elegans methods can be found at http://www.wormbook.org.

Freezing Pristionchus and other Diplogastrid nematodes- because the C. elegans freezing protocol will not work. Adapted from Nolan (1988) and requires the use of DMSO in the freshly-made freezing buffer and a thawing solution. (click here for pdf)

Making transgenic Pristionchus pacificus– it is much more difficult to obtain transgenic animals in P. pacificus than in C. elegans, but possible (1 in 250 injected)! Getting used to the ventral migration of the P. pacificus gonads and making high quality genomic DNA with compatible ends to the target transgene and the coinjection marker (Ppa-egl-20::rfp) is key to success. RNAi does not work by feeding but useful for knocking down a sensitized allele, i.e. a dominant roller or weak egl. See Cinkornpumin et al for a video feature and click here for a detailed protocol.