GSM9410254: HCT116 parental cells, biological replicate 1; Homo sapiens; RNA-Seq
Total RNA was isolated from cultured cells using TRIzol reagent following the manufacturer's instructions. RNA concentration and purity were measured with a NanoDrop 2000 spectrophotometer. RNA integrity was assessed to ensure high-quality samples for sequencing (RIN > 8.0). RNA-seq libraries were constructed using standard Illumina protocols for poly(A) selected RNA. Briefly, mRNA was purified from total RNA using oligo-dT beads, fragmented, and reverse transcribed to generate first-strand cDNA using random hexamer primers. Second-strand cDNA was synthesized, end-repaired, A-tailed, and ligated with Illumina sequencing adapters. Libraries were PCR amplified, size-selected, and quality-checked before sequencing on the Illumina NovaSeq 6000 platform with 150 bp paired-end reads.