Sequencing of 550 cancer-related genes and selected introns of human cholangiocarcinoma cell line : TFK-1
A total amount of 0.5g genomic DNA per sample was used as input material for the DNA library preparation. Sequencing libraries were generated using Agilent SureSelectXT Custom (Agilent Technologies, CA, USA) following manufacturer's recommendations and index codes were added to each sample. Briefly, fragmentation was carried out by hydrodynamic shearing system (Covaris, Massachusetts, USA) to generate 180-280bp fragments. Remaining overhangs were converted into blunt ends via exonuclease/polymerase activities and then the enzymes were removed. After adenylation of 3 ends of DNA fragments, adapter oligonucleotides were ligated. DNA fragments with ligated adapter molecules on both ends were selectively enriched by PCR reaction. After PCR reaction, library hybridizes with liquid phase with biotin labeled probe, then magnetic beads with streptomycin were used to capture the targeted regions. Captured libraries were enriched in another PCR reaction to add index tags to prepare for hybridization. Products were purified using AMPure XP system (Beckman Coulter, Beverly, USA) and quantified using the Agilent high sensitivity DNA assay on the Agilent Bioanalyzer 2100 system.