title: TL-19-F73578 NORMAL DNA
design: Tumor DNA was extracted from tumor tissue sections with tumor cellularity higher than 20% and proteinase K digestion was performed. Total nucleic acid extraction was performed with a Chemagic360 instrument using a source-specific magnetic bead protocol. Total nucleic acid was utilized for DNA library construction, while RNA was further purified by DNaseI digestion and magnetic bead purification. The nucleic acid was quantified by a Quant-iT picogreen dsDNA reagent Kit or Quant-iT Ribogreen RNA Kit (Life Technologies), and quality confirmed using a LabChip GX Touch HT Genomic DNA Reagent Kit or LabChip RNA High HT Pico Sensitivity Reagent Kit (PerkinElmer). For DNA library construction, 100 ng of DNA for each tumor and normal sample was mechanically sheared to an average size of 200 base pairs using a Covaris ultrasonicator. The libraries were prepared using the KAPA Hyper Prep Kit. Briefly, DNA underwent enzymatic end-repair and A-tailing, followed by adapter ligation, bead-based size selection, and PCR. After library preparation, each sample was hybridized to a custom designed probe set. Recovery and washing of captured targets were performed using the SeqCap hybridization and wash kit. The captured DNA targets were amplified using the KAPA HiFi HotStart ReadyMix.