The formalin-fixed paraffin-embedded (FFPE) DNA was extracted using the MagPure FFPE DNA LQ Kit following the manufacturer’s instruction. The sample’s concentration was detected by Qubit fluorometer. The integrity and purification of samples were detected by agarose gel electrophoresis. One nanogram and more FFPE DNA were randomly fragmented by Covaris. Fragmented DNAs were tested by Agilent 2100 and purified by the Agencourt AMPure XP kit. The selected fragments were end-repair and 3′ adenylated; at the same time, the adapters were ligated to the ends of these 3′ adenylated fragments. These fragments were amplified with KAPA HiFi HotStart DNA polymerase, and the PCR products were purified with the Agencourt AMPure XP kit. The library was qualified by the Agilent 2100 Bioanalyzer and ABI StepOnePlus real-time PCR (RT-PCR) system. The qualified libraries were sequenced pair end on the Hiseq4000/Xten/Novaseq system (BGI, Shenzhen, China). We used Genome Analysis Toolkit software to detect (Single Nucleotide Variants) SNVs and indels. All SNVs and indels were filtered and estimated via multiple databases, including Genome Aggregation Database and Exome Aggregation Consortium. Common variants and rare variants were classified according to minor allele frequencies (MAFs; common variants: MAF ≥0.01, rare variants: MAF <0.01).14 (link)