RNAseq for Identification of Doxycycline transcriptomic impact in N2 & CB4856 C. elegans worms

Purpose: Transcriptomic exploration for the identification of genes affected upon Doxycycline treatment in each of the two different worm strains studied Design: Wild-type C. elegans (N2 Bristol and CB4856 Hawaii) were received from Andersen’s group. Bacterial strain E. coli OP50 and E. coli HT115 were obtained from the Caenorhabditis Genetic Center (CGC). Worms of each strain were cultured on plates seeded with E. coli OP50, then eggs were obtained by alkaline hypochlorite treatment of gravid adults. A synchronized L1 population was obtained by incubating the egg suspension in M9 butter overnight at room temperature. Approximately 2000 L1 worms were transferred onto plates with or without 15 μg/mL Dox seeded with E. coli HT115. L4 worms were harvested after 50 h by three times of washing with M9 buffer. Tubes containing worm pellets were immediately submerged in liquid nitrogen for snap freezing and stored at -80ºC until use. Three biological replicates were collected for each condition.