GSM7850340: SR2; Homo sapiens; RNA-Seq
Total RNA from each sample was extracted from venous blood using the TRIzol reagent (Life Technologies, California, USA) following the manufacturer's instructions. Specific rRNA probes were used to remove rRNA from the total RNA. The remaining RNA was randomly fragmented by adding Fragmentation Buffer. Using RNA as a template, random primers were used to synthesize the first strand of cDNA. Subsequently, the RNA chain was degraded using RNase H, and the second strand of cDNA was synthesized using dNTPs as a source. The double-stranded cDNA was then purified, and end repair, A-tailing, and sequencing adapter ligation were performed. Fragment size selection was carried out using AMPure XP beads. The cDNA's second strand containing uracil (U) was degraded using USER enzyme, and the cDNA library was finally enriched through PCR.