2.6 Probe design description:
The design of liquid-phase hybridization capture probe was based on the GenCap technology of MyGenostics25. In addition to designing probes that cover the exon region of related genes (covering the exon region, exon flanks ~50bp, and non-coding disease-causing regions reported by HGMD database), encryption-designing probes were carried out for 25 CNV high-risk genes (increasing the coverage of 300bp intron region by designing probes on the exon flanks of related genes). On the other hand, designing probes also included full-length coverage ofHBA1 /HBA2 /HBB /SMN1 /SMN2 genes (coding region and intron region of coverage gene). The probes covered hot spot variation of mitochondrial genes, MT-RNR1 , MT-TL1 ,MT-ND3, and MT-ATP6 . Some regional genes of Prader-William/Angleman, DiGeorge, and Williams in microdeletion syndrome were covered, and the probe encryption was beneficial for the analysis of copy number variation in related regions. Meanwhile, according to the GC content, region size, Tm value and other parameters of the target region, the length, density, and position of the probe are adjusted based on the Probe design software BaitDesigner of MyGenostics to improve the capture efficiency of the probe.