Ion Torrent Bioinformatic analysis
Sequencing reads were aligned to hg19 reference genome. Variant calling
was performed using Ion Torrent suite software v5.12 or Ion Reporter
software v5.10. Called variants were filtered by allele frequencies,
selecting variants with MAF <1% annotated with GnomAD
database (https://gnomad.broadinstitute.org/). Nonsense, frameshift and
canonical splicing variants were considered potentially pathogenic,
while the other variants were classified according to Varsome database
(https://varsome.com/). Called variants with the minimum coverage of 20X
were analysed by Integrative Genome Viewer and confirmed by sanger
sequencing. All regions covered <20X were reanalysed by sanger
sequencing.