GSM9431135: RNA seq of EAP-3 Rattus norvegicus; Rattus norvegicus; RNA-Seq
Total RNA was extracted from rat brain tissues using the TRIzol® Reagent (Invitrogen, USA) following the manufacturer's standard protocol. The purity, integrity, and concentration of the extracted RNA were evaluated using a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, USA) and an Agilent 2100 Bioanalyzer (Agilent Technologies, USA). Only RNA samples with an RNA Integrity Number (RIN) ≥ 8.0 and A260/A280 ratio between 1.8 and 2.1 were selected for subsequent experiments. RNA-seq libraries were constructed using the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) according to the kit instructions. Briefly, poly(A)-tailed mRNA was enriched from total RNA using oligo(dT) magnetic beads, then fragmented into short segments (approximately 200 bp) under elevated temperature. First-strand cDNA was synthesized using random hexamer primers and reverse transcriptase, followed by second-strand cDNA synthesis with DNA polymerase I and RNase H. The cDNA fragments were then subjected to end repair, adenylation of 3' ends, and ligation of Illumina sequencing adapters. Finally, the ligation products were purified and amplified by PCR to generate the final cDNA libraries.