title: GSM9478889: c. elegans, young adult, agar:scaffold, rep3; Caenorhabditis elegans; RNA-Seq
library_construction_protocol: The RNA was isolated using the RNeasy Plus Universal Mini kit. Total RNA was quantified and its integrity was assessed using 5K / RNA / Charge Variant Assay LabChip and RNA Assay Reagent Kit (Perkin Elmer). Libraries were generated from 250 ng of total RNA as following: mRNA enrichment and library preparation were performed using the Illumina Stranded mRNA Prep Kit (Illumina), as per the manufacturer's recommendations. Libraries were quantified using the KAPA Library Quanitification Kits - Complete kit (Universal) (Kapa Biosystems). Average size fragment was determined using a LabChip GX II (PerkinElmer) instrument. The libraries were normalized and pooled and then denatured in 0.02N NaOH and neutralized using HT1 buffer. The pool was loaded at 200pM on a Illumina NovaSeq S4 lane using Xp protocol as per the manufacturer's recommendations. The run was performed for 2x100 cycles (paired-end mode). A phiX library was used as a control and mixed with libraries at 1% level. Base calling was performed with RTA v3. Program bcl2fastq2 v2.20 was then used to demultiplex samples and generate fastq reads.