Illumina HiSeq 2000 sequencing; Baseline gene expression profile of human extrahepatic biliary cells/biliospheres in vitro
Baseline gene expression profile of human extrahepatic biliary cells/biliospheres in vitro
Trizol lysis, chloroform extraction, aqueous layer precipitated using 70% EtOH, and immediately followed using Qiagen RNeasy mini columns.  RNA samples resuspended in Qiagen RNase-free water. NuGEN Ovation V1 kit was used for preparation of cDNA from 5ng of total RNA. For the next steps in library construction we used Epicentre End-It DNA End-Repair Kit, NEB A-Tailing kit, T4 Ligase to ligate PE Adapters and Phusion  High-Fidelity DNA polymerase for the PCR library enrichment after gel size selection.