GSM9778463: B.sub_CBS/CSE1; Drosophila melanogaster; RNA-Seq
Total RNA was isolated using RNAzol RT reagent (Molecular Research Center, USA). Briefly, 10–20 flies were homogenized in 200 μL of RNAzol reagent, after which 80 μL of dH₂O was added, and the mixture was incubated on ice for 10 minutes. The homogenate was then centrifuged at 15,000×g for 10 minutes at 4°C. The resulting supernatant was collected and transferred to a new sterile 1.5-mL tube. An additional purification step was performed by adding 1 μL of 4-bromoanisole (MRC, BN 191). The mixture was incubated at room temperature for 3 minutes and centrifuged again at 15,000×g for 10 minutes at +4°C. The supernatant was transferred to a new sterile tube, and 200 μL of isopropyl alcohol was added to precipitate the RNA. The mixture was incubated on ice for 30 min and then centrifuged at 20,000×g for 20 min at +4°C. The RNA pellet was washed twice with chilled 75% ethanol and dissolved in RNase-free water. RNA-seq libraries were prepared using the VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina (cat# NR605-01, Vazyme Biotech, China) according to the manufacturer's protocol. Sequencing was performed on an Illumina NextSeq 500 platform