GSM8996249: Control-3; Drosophila melanogaster; RNA-Seq
After 24 hours of starvation, the abdomens of adult female flies were dissected on ice and immediately transferred into TRIzol reagent. Tissues were homogenized thoroughly, and total RNA was extracted following the standard TRIzol protocol, including chloroform phase separation, isopropanol precipitation, ethanol washing, and resuspension in RNase-free water. RNA quality and concentration were assessed using a Nanodrop or Qubit before sequencing. Total RNA was treated with DNase I to remove genomic DNA contamination, and mRNA was enriched using poly(A) selection. The purified mRNA was fragmented and reverse transcribed into cDNA using random primers. After second-strand synthesis, the cDNA was end-repaired, A-tailed, and ligated to sequencing adapters. The ligated products were PCR-amplified to generate the final sequencing libraries, which were quantified, assessed for quality, and then subjected to high-throughput sequencing.