Figure 4. Sanger sequencing validation of duplication breakpoints at 6q16.2. The proband (II-1) and all available affected relatives (Appendix 1) were tested with primers designed across the predicted breakpoints to generate a unique junction fragment sequence. The exact breakpoint is marked with black vertical arrows; the location of polymerase chain reaction (PCR) primers is represented with red arrows. Genomic coordinates (Hg38) indicating the end of the first copy and the start of the second copy are indicated. An additional 15-bp sequence is inserted into the breakpoint of the duplicated sequence.