GSM8068172
Total RNA was extracted from the cells using ISOGEN II (Nippon Gene). Total RNA samples were first enriched for mRNA with poly A tails using oligo dT beads, then fragmented into small pieces, and synthesized into first strand cDNA using random primers. The second strand cDNA was synthesized using dUTP instead of dTTP, followed by end-repair and 3' adenylation of the synthesized cDNA, with adaptors ligated to these ends. The U-labeled second-strand template was digested with Uracil-DNA-Glycosylase (UDG) and amplified by PCR, after which the library underwent quality control, circularization, and amplification to create DNA nanoballs (DNB), which were then sequenced using the DNBSEQ Technology platform.