ASXL3, p.R1004Efs*21 - Sanger sequencing verification of NGS finding in proband and her parents

The PCR were performed using PCR kit by Promega. Termination reactions were performed using BigDye v3.1 termination kit (ThermoFischer Scientific).

DNA primers were designed using online tools Primer3, Primer Blast, UCSC In-Silico PCR (5’-CAGAGCAACACAGCTTTGGA-3‘ as forward primer and 5‘-GGAGACATTTCCAGGCCCTAT-3 as reverse primer).Before any analyses, the proband's parents signed the informed consent. The analyses were performed as laboratory tests at the Department of Medical Genetics, University Hospital Brno (Czech Republic).