title: GSM9372924: 40day_w1118_head_1; Drosophila melanogaster; RNA-Seq
library_construction_protocol: Following collection at ZT8 from age-appropriate flies, head and gut samples were dissected in 1X PBS, homogenized in TRIzol Reagent (Vazyme, Cat# R401-01), and subsequently processed for RNA extraction and RNA-seq analysis. Following quality control, eukaryotic mRNA was enriched using oligo(dT) beads that bind to the poly-A tail via A-T complementarity. The mRNA was then fragmented, and first-strand cDNA was synthesized using random hexamers. Second-strand cDNA was generated using buffer, dNTPs, and DNA polymerase I, followed by purification with AMPure XP beads. The purified double-stranded cDNA underwent end repair, A-tailing, and sequencing adapter ligation. Fragments of appropriate size were selected again with AMPure XP beads, and the final cDNA library was amplified via PCR.