DNA-Seq gene panel of a human patient with metastatic RCC: tumor sample 2
DNA was extracted from fresh-frozen tumoral tissue after pathological selection using a Maxwell 16 MDx Instrument (Promega, Promega Biotech Ibrica S.L., Madrid, Spain) and quantified by Qubit (Qiagen, Madrid, Spain). We sequenced sequential paired samples from the same patient in an Ion S5 next-generation sequencing (NGS) system using the Ion AmpliSeq Comprehensive Cancer Panel (ThermoFisher Scientific, Madrid, Spain), which targets the exons of 409 tumor suppressor-genes and oncogenes. In brief, 25 ng tumor DNA per primer pool for a total of 100 ng input DNA were used for sequencing. The Ion AmpliSeq Comprehensive Cancer Panel is optimized for library construction with the Ion AmpliSeq Library Kit 2.0. Libraries are then ready for template preparation on the Ion OneTouch System and sequencing on the Ion S5 Sequencer.