Blood genome column medium extraction kit

A peripheral blood sample was obtained from the patient in an EDTA anticoagulant blood sample tube that was stored at 4°C for less than 6 h. DNA was extracted using the Blood Genome Column Medium Extraction Kit (Tiangen Biotech, Beijing, China) according to the manufacturer's instructions. Protein‐coding exome enrichment was performed using the xGen Exome Research Panel v1.0, comprising 429,826 individually synthesized and quality‐controlled probes targeting 49.11 Mb of protein‐coding region (>23,000 genes) of the human genome. WES was performed using the NovaSeq 6000 platform (Illumina, San Diego, CA, USA), and the raw data were processed using FastP to remove adapters and filter low‐quality reads. Paired‐end reads were aligned to the Ensembl GRCh38/hg38 reference genome using the Burrows–Wheeler Aligner. Variant annotation was performed in accordance with database‐sourced minor allele frequencies (MAFs) and practical guidelines on pathogenicity issued by the American College of Medical Genetics. The annotation of MAFs was performed based on the 1000 Genomes, dbSNP, ESP, ExAC, and Chigene in‐house MAF database, Provean, Sift, Polypen2_hdiv, and Polypen2_hvar databases using R software (R Foundation for Statistical Computing, Vienna, Austria).

A peripheral blood sample was obtained from the patient in an ethylenediaminetetraacetic acid (EDTA) anticoagulant blood sample tube that stored at 4°C for less than 6 hr. DNA was extracted using the Blood Genome Column Medium Extraction Kit (Tiangen Biotech, Beijing, China) according to the manufacturer's instructions. Protein‐coding exome enrichment was performed using the xGen Exome Research Panel v1.0, comprising 429,826 individually synthesized and quality‐controlled probes targeting 49.11 Mb of protein‐coding region (>23,000 genes) of the human genome. WES was performed using the NovaSeq 6000 platform (Illumina, San Diego, CA, USA), and the raw data were processed using FastP to remove adapters and filter low‐quality reads. Paired‐end reads were aligned to the Ensembl GRCh38/hg38 reference genome using the Burrows–Wheeler Aligner. Variant annotation was performed in accordance with database‐sourced minor allele frequencies (MAFs) and practical guidelines on pathogenicity issued by the American College of Medical Genetics. The annotation of MAFs was performed based on the 1000 Genomes, dbSNP, ESP, ExAC, and Chigene in‐house MAF database, Provean, Sift, Polypen2_hdiv, and Polypen2_hvar databases using R software (R Foundation for Statistical Computing, Vienna, Austria). The sequencing data have been deposited in GSA database (http://ngdc.cncb.ac.cn/gsub/) (HRA001665).

Peripheral blood samples were obtained from the patients in an EDTA anticoagulant blood sample tube stored at 4°C for less than 6 h. DNA was extracted using the Blood Genome Column Medium Extraction Kit (Tiangen Biotech, Beijing, China) in accordance with the manufacturer's instructions. Protein‐coding exome enrichment was performed using the xGen Exome Research Panel v.1.0, comprising protein‐coding regions (>23,000 genes) of the human genome. Exome sequencing was performed using the NovaSeq 6000 platform (Illumina, San Diego, CA, USA), and the raw data were processed using FastP to remove adapters and filter out low‐quality reads. Paired‐end reads were aligned to the ENSEMBL GRCh38/hg38 reference genome using the Burrows–Wheeler Aligner. Variant annotation was performed in accordance with database‐sourced minor allele frequencies (MAFs) and practical guidelines on pathogenicity issued by the American College of Medical Genetics. The annotation of MAFs was performed based on the 1000 Genomes, dbSNP, ESP, ExAC, Provean, Sift, Polypen2_hdiv, Polypen2_hvar, and Chigene in‐house MAF databases using R software (R Foundation for Statistical Computing, Vienna, Austria). The sequencing data have been deposited in the GSA database (http://ngdc.cncb.ac.cn/gsub/) (HRA001920).