GSM9149382: SCC-9,sh-ZIC2,rep3; Homo sapiens; RNA-Seq
RNA was extracted using the NEBNext Ultra II RNA Library Prep Kit for Illumina, and a total RNA amount of ≥1 μg was selected for sequencing library construction. mRNA with polyA tails was enriched using Oligo(dT) beads and then randomly fragmented. The fragmented mRNA was used as a template for cDNA synthesis with random oligonucleotide primers. The synthesized cDNA was purified, followed by end repair, addition of a single "A" nucleotide at the 3' ends, and ligation of sequencing adapters. cDNA fragments of approximately 400–500 bp were selected using AMPure XP beads. The selected fragments were then PCR-amplified and purified again using AMPure XP beads to generate the final sequencing library.