GSM8546139: Cardiomyocytes2, DMSO, 24h; Rattus norvegicus; RNA-Seq
RNA was harbested using Dynabeads Oligo (dT)25-61002 (ThermoFisher). Fragmentation buffer was added to break the mRNA into short segments. Using the mRNA as a template, first-strand cDNA was synthesized with random hexamer primers. Following this, a buffer, dNTPs, DNA polymerase I, and RNase H were added to synthesize double-stranded cDNA, which was then purified using AMPure XP beads.  The purified double-stranded cDNA underwent end repair, A-tailing, and ligation of sequencing adapters, followed by size selection using AMPure XP beads. PCR amplification was then performed, and the PCR products were purified with AMPure XP beads to obtain the final library. Once the library construction was complete, preliminary quantification was conducted using a Qubit 2.0, and the library was diluted to 1.5 ng/µL. The insert size of the library was assessed using an Agilent 2100. After confirming that the insert size met expectations, accurate quantification of the library's effective concentration was performed using Q-PCR (with effective concentration > 2 nM) to ensure library quality.