WXS of human : children blood
1.Method1.1 Blood samples were collected from the patient, and genomic DNA was extracted with QIAamp DNA Blood Mini kit (Qiagen, Hilden, Germany) following the manufacturers protocol.Informed consent was obtained from the patient and her parents.1.2 Next-generation sequnencingThe extracted DNA was fragmented randomly and then purified using magnetic particle method.DNA fragments were ligated with adaptors and captured by probes of the IDT XGen Exome Research Panel (IDT, Lowa, USA) targeting about 19,396 genes.The DNA libraries after enrichment and purification were sequenced on the NovaSeq 6000 sequencer according to the manufacturers instructions (Illumina, San Diego, USA).1.3 Data analysisAll reads were aligned to the reference human genome (UCSC hg19) by Burrows-Wheeler Aligner (BWA)(v.0.5.9-r16).[1] After data annotation using PriVar toolkit,clinical significance of the variants were identified.The detected variants were confirmed using PCR and the products were subjected to direct sequencing on 3500XL Genetic Analyzer(Applied Biosystems, Foster City, USA) according to the manufacturers instructions.