GSM8384017: N2 Celegans late stage embryos-replicate2; Caenorhabditis elegans; RNA-Seq
C. elegans RNA was extracted as described (Green and Sambrook, 2020). Briefly, late stage embryos were collected from worms grown on five NGM plates and flash frozen in liquid N2. The frozen pellet was grinded to a fine powder and dissolved in ten volumes of TRIzol™ (Invitrogen, USA). Total RNA was isolated according to the manufacturer's protocol, flash frozen in liquid N2 and stored at -80 °C. For RNA-seq, library preparation and sequencing were performed by Novogene (Novogene Co, Hong Kong). Poly-T oligo-attached magnetic beads (NEB Oligo d(T)25 Magnetic Beads, S1419S) were used to purify mRNA from total RNA. Random hexamer primers were used to synthesize first-strand cDNA (included in the library preparation kit), followed by second-strand cDNA synthesis using dUTP for strand-specific libraries. NEBNext® Ultra™II Directional RNA Library Prep Kit(Cat No. E7760) was used for library preparation. Qubit (Thermo fisher scientific) and real-time PCR were used for quantification and quality control. The size distribution of the library was analyzed using Agilent bioanalyzer (2100). Quantified libraries were pooled and sequenced on Illumina NovaSeq 6000 PE150 platform.