title: GSM3382019: Embryo 6-11 hrs S5; Tribolium castaneum; RNA-Seq
library_construction_protocol: Embryos were collected at 0-5 hours and 6-11 hours respectively in three replicates (each replicate has 80-100 Embryos). Similarly, ovary, testis and carcasses from adult beetles at four days post eclosion (30 beetles in each replicate) were collected to evaluate transcriptional activity. Total RNA from these tissues was extracted with Trizol (Invitrogen, Carlsbad, CA, USA) following the manufacturer's instructions. After total RNA extraction, RNA was treated with Turbo DNase (Ambion/Applied Biosystems, Austin, TX). RNAseq library was prepared using NEBNext Poly (A) mRNA Magnetic Isolation Module (NEB #E7490) for mRNA isolation and NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB #E7530S) for library preparation according to the manufacturing instructions. Each library had a peak size of approximately 300 base pairs. cDNA library quality was assessed on an Agilent 2200 TapeStation using a High Sensitivity D1000 tape. Libraries were quantified using the Invitrogen Qubit 2.0 Fluorometer High Sensitivity dsDNA assay. All libraries were normalized to a 4 nM concentration and then pooled in equal volumes for denaturing and diluting for sequencing. The library pool was denatured and diluted to a final concentration of 1.8 pM following the Illumina NextSeq Denature and Dilution protocol including a 1% PhiX DNA spike in
identifiers.namespace: GEO
identifiers.id: GSM3382019