GSM9464490: S66_adK; Homo sapiens; RNA-Seq
48h post adenoviral infection, media was removed, and each well was washed with 1X cold PBS. RNA was extracted using 500uL of Trizol (Invitrogen, Carlsbad, CA, USA) following manufacturer's instructions. For protein extraction, fibroblasts were culture for 72h post adenoviral infection. Conditioned media was saved, and each well was washed with 1X cold PBS. Lysates were harvested by scraping in 200uL of 2X SSB (125mM Tris pH 6.8, 10% glycerol, 3% SDS, 0.01% bromophenol blue, 715mM beta-mercaptoethanol). mRNA library preparation with poly A enrichment/non-directional. 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 dTTP for non strand specific library or dUTP for strand specific library.