DNA and RNA extraction and quantification
Germline DNA and RNA were extracted from peripheral blood using the MagNA Pure Compact extractor system (Roche Diagnostics), according to the manufacturer’s recommendations. The PAXgene Blood RNA Kit (PreAnalytiX) was used to extract germline RNA when the patient could not come to our hospital. Tumor DNA and RNA were extracted from 7µm-thick FFPE tissue sections, and a hematoxylin and eosin-stained section of each block allowed the assessment of tumor cell area and content by two experienced pathologists. Tumor DNA was purified using the QIAamp DNA FFPE Tissue Kit from Qiagen, while tumor RNA was isolated employing the RNeasy FFPE Kit (Qiagen), according to their corresponding protocols. The NanoDrop ND-1000 Spectrophotometer was used to assess the DNA and RNA quantity and quality. However, for NGS purposes, the quality of DNA samples was also tested by agarose gel electrophoresis and the concentration of the samples was measured in a Qubit 3.0 Fluorometer (Life Technologies) and a 2100 Bioanalyzer (Agilent).