DNA extraction and library preparation
tumor
TUMOR
A minimum of 20% tumor content was required needed for clinical formalin-fixed paraffin-embedded (FFPE) samples used for genomic evaluation. Genomic DNA (gDNA) of formalin-fixed and parrffin-embedded (FFPE) tissues was extracted using Genomic DNA Tissue Extraction Kit (Concert®),and matched peripheral blood was extracted using Magnetic Universal Genomic DNA Kit (TIANGEN). NGS tests targeting Whole exome Hyb Panel were performed at Simceredx company (Nanjing,China), and following manufacturers’ instructions. In brief, 200 ng gDNA was sheared into 200~300 bp by enzymatic fragmentation kit. Indexed paired-end adaptors for Illumina platform were synthesized by Integrated DNA Technologies (IDT). End repair, A-tailing, and adaptor ligation of sheared DNA were performed with the reagents from KAPA Hyper DNA Library Prep kit (Roche Diagnostics). Unligated adaptors were removed by the size selection function of Agencourt AMPure XP beads (Beckman Coulter) and the ligation products were PCR amplified to form a prelibrary for hybridization.