title: Illumina HiSeq 4000 sequencing; Effect of Cadmiun and Perfluorooctanoic acid (PFOAs) chemicals on global gene expression in liver cells
design: Effect of Cadmiun and Perfluorooctanoic acid (PFOAs) chemicals on global gene expression in liver cells
library_construction_protocol: HepaRG and HepaG2 cells treated with physiological concentrations of Cadmiun or PFOAs for 2 months in triplicate and samples were collected.  Total RNA was isolated using TRIZOl reagents (Sigma). RNA sequencing libraries were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina following manufacturer's instructions (NEB, Ipswich, MA, USA). Briefly, mRNAs were first enriched with Oligo(dT) beads. Enriched mRNAs were fragmented for 15 minutes at 94 °C. First strand and second strand cDNAs were subsequently synthesized. cDNA fragments were end repaired and adenylated at 3'ends, and universal adapters were ligated to cDNA fragments, followed by index addition and library enrichment by limited-cycle PCR. The sequencing libraries were validated on the Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified by using Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA) as well as by quantitative PCR (KAPA Biosystems, Wilmington, MA, USA).