GSM9302239: C4-2 cells, 25 mM glucose, biological replicate 1; Homo sapiens; RNA-Seq
RNA was harvested using Direct-zol RNA Miniprep Kit (Zymo Research). Total RNA >= 1 µg was used for construction of sequencing libraries. Poly(A)+ mRNA was enriched from total RNA using Oligo(dT) magnetic beads. The mRNA was fragmented with fragmentation buffer, and first-strand cDNA was synthesized using random hexamer primers. Second-strand cDNA was synthesized using DNA polymerase I and dNTPs, followed by purification with magnetic beads. Double-stranded cDNA underwent end repair, A-tailing, and adaptor ligation. After fragment size selection and PCR enrichment, the final libraries were quantified and assessed for insert size. Libraries passing QC were sequenced on an Illumina platform