Transcriptome analysis in C.elegans from Parental P0 to F3 generations under hypoxic stress in both WT and hrde-1 mutant [RNA-seq]

The goal of RNA-seq is to identify the differentially expressed genes in the wild-type (WT) and hrde-1 mutant of C.elegans at the normoxia reared and hypoxia treatment conditions at P0 generation, and those maintained dysregulated genes in F1, F2 and F3 generation. Two biological replicates were assigned for each group and in total 32 groups were prepared for RNA-seq libraries with 500 ng mRNA as starting materials using NEXTflex Illumina qRNA-Seq Library Prep Kit (Bioo Scientific). We mapped about 20 - 30 million reads per sample to C.elegans (WS235) with RSEM pipline. Differential analysis were conducted using EdgeR. The significant genes were conducted by computing NB exact genewise tests with corrected p < 0.05. Overall design: Messenger RNA from WT and hrde-1 under normoxia or hypoxia treatment worms from P0 to F3 generations were extacted, and cDNA libraries were generated for deep sequencing, in duplicate, using paired-end 150bp on Illumina Hiseq4000 platform