4T1_shCrabp1-1 (CRX2193164)
Total RNA was isolated, followed by poly(A)+ mRNA enrichment using oligo(dT) magnetic beads. The purified mRNA was fragmented under optimized conditions, and double-stranded cDNA was synthesized via random hexamer-primed reverse transcription. Subsequent library preparation steps included end repair, 3'-end A-tailing, and ligation of Illumina sequencing adapters. Libraries were then size-selected using AMPure XP beads (Beckman Coulter) and quantified via high-sensitivity assays (Qubit fluorometry and Bioanalyzer electrophoresis) to ensure optimal sequencing quality.