mRNA-seq of wildtype C. elegans L3 larvae sampled at molt entry and molt exit [RNAseq_MoltEntry-MoltExit_N2]

In order to determine the gene expression profile of worms at molt entry and exit in more detail, we collected animals specifically at these points based on signal from a luciferase-based developmental assay (Olmedo et al., 2015, Genetics). This approach allowed us to pool sufficient animals to perform mRNA sequencing in order to specifically determine the transcriptome of animals at these developmental stages. Using these transcriptome profiles,we can re-align time course RNA-seq datasets to the beginning of a larval stage (molt exit). Synchronized L1 animals, expressing luciferase, were grown while measuring developmental trajectories using a luminometer. Assay was stopped when the majority of animals were at L3 molt entry and L3 molt exit respectively. Based on luciferase traces, individual animals of the appropriate developmental timepoint were collected from their wells and pooled for RNA isolation and library preparation.