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.