cDNA-Seq of LZX: adult male blood
materials and methods: Whole-exome sequencing (WES) was performed on the probands. Genomic DNA was extracted from whole blood samples using the QIAamp DNA Blood Mini Kit (Qiagen Inc., USA) following the manufacturers protocol. The quantity and quality of obtained DNA samples were analyzed by NanoDrop 2000 (Thermo Scientific, USA) and Agilent 2100 bioanalyzer (Agilent Technologies, USA). Agilent SureSelect Human All Exon V6 (Agilent Technologies, USA) was used to enrich DNA fragments of human exons after the fragmentation of genomic DNA. Sequencing data was produced on Illumina NovaSeq platform. The adaptor sequences were trimmed from the tail of sequencing reads using cutadapt (v1.15). Sequencing reads were aligned to human reference genome (hg19) with BWA (v0.7.15). Duplicated reads were marked by Picard (v2.4.1). Qualimap (v2.2.1) was used to calculate base quality metrics, genome mapping rate, and the coverage of targeted regions. Base quality score recalibration, single nucleotide variants (SNVs) and small insertions or deletions (InDels) calling were performed following the best practice protocol of GATK (v3.8). Variant filtering was done by a finely tuned in-house script. Pass-filter variants were annotated using VEP (release 88).