WXS of Pat101's lymphnode
Genomic DNA was extracted from the FFPE tumor and paired lymph node tissues using a High Pure FFPET DNA Isolation Kit (Roche, Switzerland) according to the manufacturer's protocol. DNA isolation from peripheral blood was performed with a MagNA Pure Compact Nucleic Acid Isolation Kit I (Roche) on a MagNA Pure Compact Instrument (Roche). A Qubit 4.0 Fluorometer (Thermo Fisher Scientific, USA) was used to measure DNA concentration. DNA quality was assessed by quantitative PCR (qPCR) using QuantumDNA Kit (Evrogen, Russia). Isolated total DNA (100 ng) was used to construct sequencing libraries with Rapid Capture Exome Kit (Illumina, USA) or TruSeq Exome Library Prep Kit (Illumina) following manufacturers protocols. Sequencing of exome libraries was performed on a NextSeq500 System (Illumina) using a 75 or 150 paired-end (PE) sequencing strategy.