RNA sequences of AC16 cardiomyocytes from human samples.
After agarose gel electrophoresis and quality assessment and quantification using the Nanodrop spectrophotometer, the total RNA samples were subjected to mRNA enrichment using oligo(dT) magnetic beads. In the case of degraded RNA samples or samples derived from prokaryotes, an rRNA removal kit was employed for the enrichment step. The RNA sequencing libraries were constructed using commercially available kits. This involved RNA fragmentation, followed by random primer reverse transcription to generate the first-strand cDNA. The second-strand cDNA synthesis incorporated dUTP for subsequent strand-specific library preparation. The resulting double-stranded cDNA underwent end repair and A-tailing, followed by ligation of Illumina adapter sequences. Finally, PCR amplification was performed to obtain the final libraries. The quality of the constructed libraries was assessed using an Agilent 2100 Bioanalyzer, and library quantification was conducted using qPCR. The libraries were sequenced using the Illumina NovaSeq 6000 platform.