GSM9648621: HT22, transfected, 15min, incubated, 24h, rep1; Mus musculus; miRNA-Seq
Total RNA was extracted from the cells using TRIzol reagent. After the samples passed quality control, eukaryotic mRNA was enriched using Oligo(dT)-conjugated magnetic beads. The enriched mRNA was then randomly fragmented by adding fragmentation buffer. Using the fragmented mRNA as templates, first‑strand cDNA was synthesized with random hexamers, followed by second‑strand cDNA synthesis using buffer, dNTPs, and DNA polymerase I. The double‑stranded cDNA was purified with AMPure XP beads. The purified double‑stranded cDNA was then subjected to end repair, A‑tailing, and adapter ligation. Fragment size selection was performed using AMPure XP beads, and finally, PCR amplification was carried out to obtain the final cDNA library.