GSM9499692: Transcriptome of RKO cell (DMSO control), rep 1; Homo sapiens; RNA-Seq
Total RNA was isolated from RKO cells using TRIzol reagent according to the manufacturer's instructions. RNA integrity was verified by agarose gel electrophoresis. RNA purity and concentration were assessed using a NanoDrop spectrophotometer, with acceptance criteria of OD260/280 ≥ 1.8 and OD260/230 ≥ 2.0. Sequencing libraries were constructed from 1 µg of high-quality total RNA per sample. Messenger RNA was enriched using oligo(dT) magnetic beads. The NEBNext Ultra RNA Library Prep Kit for Illumina was used for library preparation following the manufacturer's protocol, which includes mRNA fragmentation, first and second strand cDNA synthesis, end repair, dA-tailing, adapter ligation, and PCR amplification. The final cDNA libraries were size-selected for fragments of 250-300 bp and their quality was assessed.