GSM9891795: HK-2_sg_replicate2, target gene sgRNA knockdown; Homo sapiens; RNA-Seq
Total RNA was isolated from cultured HK-2 cells using Trizol reagent following the manufacturer's instructions. RNA integrity was verified, and RNA purity and concentration were determined by spectrophotometry. mRNA was enriched from total RNA using oligo (dT) magnetic beads. Fragmentation buffer was used to interrupt mRNA into short fragments. First-strand cDNA was synthesized using random hexamer primers, followed by second-strand cDNA synthesis. After end repair, A-tailing and adapter ligation, fragments were purified and enriched by PCR amplification to construct the cDNA library. The libraries were sequenced on Illumina NovaSeq 6000 platform with paired-end 150 bp strategy.