GSM5902820: CR43432_mut_8_12_mRNA_line1_sequence; Drosophila melanogaster; RNA-Seq
RNA-seq libraries were prepared using the NEXTflex Rapid Directional RNA-seq Kit (Bioo Scientific) following enrichment of polyadenylated mRNAs with NEXTflex Poly(A) Beads (Bioo Scientific). Libraries were sequenced on an Illumina HiSeq platform with 50-nt single-end reads. RNA was extracted with TRI Reagent (Life Technologies), according to the manufacturer's protocol. Briefly, samples were homogenized in TRI reagent, and RNA was phase-separated by the addition of chloroform (J.T. Baker Analytical) at a ratio of 250 µL per 1 mL TRI Reagent. RNA was then precipitated with isopropanol, with GlycoBlue (Life Technologies) added as a co-precipitant when necessary, and pellets were washed twice with 70% ethanol prior to resuspension in water.