Agilent genomic workbench 7.0 program

The CMA was performed using oligonucleotide 8x60K CytoScan^® gene chip (Agilent customer design ID 040427, Changhua Christian Hospital, Changhua, Taiwan). DNA labeling and hybridization were carried out according to manufacturer’s recommendation. Scanned images were analyzed by Feature Extraction 9.5.3 software (Agilent Technologies, Santa Clara, CA, USA), and the extracted data were processed using the Agilent Genomic Workbench 7.0 program (Agilent Technologies, Santa Clara, CA, USA). In Families 1–4, the maternal grandparents of the fetuses were enrolled for CMA. In Family 3, the maternal uncle and the maternal granduncle were also included into analyses (Figure 1). The CMA findings were described based on the reference genome version of GRCh37. Online publicly available databases used for evaluation of the clinical significance of CNVs include DECIPHER (https://decipher.sanger.ac.uk/), NCBI dbVar’s nstd102 (Clinical Structural Variants) (http:// https://www.ncbi.nlm.nih.gov/dbvar/studies/nstd102/), European Cytogeneticists Association Register of Unbalanced Chromosome Aberrations (ECARUCA) (http://www.ecaruca.net), Online Mendelian Inheritance in Man (OMIM) (http://https://omim.org/), a genome database at the University of California, Santa Cruz (UCSC) (https://genome.ucsc.edu/) and Database of Genomic Variants (DGV) (http://dgv.tcag.ca/).

Copy number analyses were performed by CMA using an Agilent customer design oligonucleotide 8 × 60 K CytoScan^® gene chip (ID 040427). DNA labeling and hybridization were carried out according to manufacturer’s recommendation. Scanned images were analyzed by Feature Extraction 9.5.3 software (Agilent Technologies, Santa Clara, CA, USA), and the extracted data were processed using the Agilent Genomic Workbench 7.0 program (Agilent Technologies, CA, USA). The CMA findings were described based on the reference genome version of GRCh37, following the latest guideline of An International System for Human Cytogenomic Nomenclature (ISCN2020).