title: GSM6924251: N2 RNA-seq, replicate 1; Caenorhabditis elegans; RNA-Seq
library_construction_protocol: TRI Reagent (Sigma Aldrich) was added to the embryo pellet followed by 1-3 cycles of snap-freezing, thawing, and vortexing (1 minute). Following chloroform extraction, the aqueous phase was transferred to an RNA Clean and Concentrator-5 column and purified according to the manufacturer's instructions including on-column DNaseI digestion (Zymo Research). PolyA+ RNA was isolated from total RNA using the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB E7490) according to the manufacturer's instructions. Libraries were prepared with the NEBNext Ultra II Directional RNA Library Prep with Sample Purification Beads (NEB E7765S) according to the manufacturer's directions with NEBNext Multiplex Oligos for Illumina (NEB E6440). Library concentration was quantified using NEBNext Library Quant Kit for Illumina (NEB E7630). Pooled libraries were sequenced using NextSeq500 in 75 bp paired-end mode.