title: GSM9003620: FB,GLU,rep9; Drosophila melanogaster; RNA-Seq
library_construction_protocol: Twenty-eight days after eclosion, adult female flies from GLU-HGLU and GLUGAL-HGLU groups were anesthetized under CO2 and dissected in cold, RNase-free PBS under a stereomicroscope, and the abdominal carcasses were isolated (the dorsal body wall tissue with removal of internal organs like digestive tract and ovary, etc., retaining primarily metabolic organs including fat body and oenocyte) and were snap frozen in liquid nitrogen. Each group had 12 biological replicates (pools of 10 abdominal carcasses per replicate). Total RNA was extracted from pooled abdomens using Invitrogen Trizol reagent (Thermo Fisher Scientific, Schwerte, Germany), according to the manufacturer's instructions. RNA integrity was verified on the Agilent 2100 Tapestation (Agilent Technologies Inc, Santa Clara, CA, USA); all samples had a RNA integrity number above 7. 1. Take a certain amount of total RNA samples, and use oligo dT beads to enrich mRNA with poly A tail; 2. mRNA molecules were fragmented into small pieces; 3. The fragmented mRNA was synthesized into first strand cDNA using random primers; 4. The second strand cDNA was synthesized with dUTP instead of dTTP; 5. The synthesized cDNA was subjected to end-repair and 3' adenylated. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments; 6. Perform PCR amplification; 7. Library quality control; 8. Library circularization; 9. The library was amplified to make DNA nanoball (DNB); 10. Sequencing on DNBSEQ (DNBSEQ Technology) platform.