GSM9154938: Female 5×FAD Rapamycin 4; Mus musculus; RNA-Seq
Total RNA was isolated from mouse cortical tissues using TRIzol reagent (Cat. #15596026, Thermo Fisher Scientific) following homogenization and lysis. cDNA fragments of 150–200 bp were size-selected using the AMPure XP system (Beckman Coulter, Beverly, USA). Adapter-ligated cDNA was treated with 3 µL of USER enzyme (NEB, USA) at 37°C for 15 min, followed by incubation at 95°C for 5 min prior to PCR amplification. PCR was performed using Phusion High-Fidelity DNA polymerase, universal PCR primers, and index (X) primers. The resulting PCR products were purified using the AMPure XP system, and library quality was assessed using the Agilent Bioanalyzer 2100 system. After passing quality control, libraries were pooled based on effective concentration and sequencing requirements, followed by paired-end sequencing (150 bp) on the Illumina HiSeq 2500 platform (Illumina, San Diego, USA) with unique index barcodes for sample identification.