Analysis of gene expression and polysome profiling in the presence and absence of glh-1 in C. Elegans

This experiment compares gene expression and polysome profiling between a strain where glh-1 is present - DUP64 glh-1(sam24[glh-1::gfp::3xFlag]) I and a strain where the glh-1 coding sequence has been deleted DUP144 glh-1(sam65[Δglh-1::gfp::3xFlag]) I For each of three replicates, 25 plates of each strain were used to inoculate 1L of S Media containing 5 grams of freeze-dried OP50 (LabTie, Leiden, The Netherlands). Liquid cultures were incubated in 4 1L beveled flasks at 20° C until worms were gravid, at which point embryos were harvested, washed, bleach-treated (Montserrat et al. 2012) and hatched overnight. The synchronized L1-staged worms were used to inoculate another 1L of S Media with OP50 and were grown for approximately 40 hours until the majority reached the young adult stage, where they were harvested, washed, pelleted and flash frozen in 1-ml aliquots.