GSM8326115: A549_WT_VitC2; Homo sapiens; RNA-Seq
Total RNA was extracted using Trizol reagent kit (Invitrogen, Carlsbad, CA,USA) according  to the manufacturer's protocol. RNA quality was assessed on an Agilent 2100 Bioanalyzer  (Agilent Technologies, Palo Alto, CA, USA) and checked using RNase free agarose gel  electrophoresis. After total RNA was extracted, eukaryotic mRNA was enriched by  Oligo(dT) beads(pronucleus:After total RNA was extracted, prokaryotic mRNA was enriched by  removing rRNA by Ribo-ZeroTM Magnetic Kit (Epicentre, Madison, WI, USA).Then the  enriched mRNA was fragmented into short fragments using fragmentation buffer and reversly  transcribed into cDNA by using NEBNext Ultra RNA Library Prep Kit for Illumina(NEB  #7530,New England Biolabs, Ipswich, MA, USA).The purified double-stranded cDNA  fragments were end repaired, A base added, and ligated to Illumina sequencing adapters.The  ligation reaction was purified with the AMPure XP Beads(1.0X).And polymerase chain  reaction (PCR) amplified.The resulting cDNA library was sequenced using Illumina  Novaseq6000 by Gene Denovo Biotechnology Co. (Guangzhou, China).