Whole exome sequencing analysis

Whole exome sequencing analysis was applied to 50 ng of genomic DNA from TIMM50 Pt. Nextera Rapid Exome Capture Kit (Illumina) was used for exome enrichment according to manufacturer's instructions. Exome sequencing was performed by Illumina MiSeq next‐generation sequencing platform with 600‐bp pair‐end sequencing strategy. Sequences from the FASTAQ files were aligned to the human genome (hg19) by using the BWA aligner. Variants were called by using the GATK HaplotypeCaller and then filtered by using the Variant Quality Score Re‐calibration according to the best practices of GATK‐2.7 (https://www.broadinstitute.org/gatk/). Variants were annotated by using ANNOVAR. Coverage of the targeted regions was estimated using the GATK DepthOfCoverage. For paired‐end reads to be included, they needed to have a mapping quality >20 and a base quality >10.