Bulk RNA-Seq of homo sapiens: PDAC cells from co-culture with Cancer Associated Fibroblasts
Cells that were cocultured were sorted by FACS and immediately underwent RNA extraction and sent to Novogene Corporation Inc for sequencing. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using either dUTP for directional library or dTTP for non-directional library. For the non-directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. For the directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification.