Genomic Profiling of Hereditary Conditions

Genomic DNA was extracted from an EDTA-treated sample of the patients’ peripheral blood using the Blood Genomic DNA Extraction Kit (Qiagen, Germany). Apolipoprotein E (APOE) genotypes were determined by multiplex amplification refractory mutation system polymerase chain reaction (Jiang et al., 2018 (link)). The DNA sample of the proband was processed using WES. The details of the WES protocols and bioinformatic analysis have been described in previous publications (Li et al., 2017 (link); Jiang et al., 2019 (link)). In brief, an enriched DNA sample was sequenced on the Illumina Nova 6000 platform (Kangso Medical Inspection Co. Ltd., Beijing, China). All the variants were annotated using ANNOVAR software. The frequency of the variants in the general population was identified by the 1000 Genomes Project¹, the genome aggregation database², the ExAC database³, and the single-nucleotide polymorphism database⁴. SIFT⁵, CADD⁶, and PolyPhen-2⁷ were used to predict the possible functional changes caused by the variants. The potential variants were verified by Sanger sequencing. All the available family members, as well as the 500 healthy individuals, were sequenced to validate the confirmed variants.