Because the transcription factor gene, ZIC2, lies close to the breakpoint and because mutations in ZIC2 are known to cause holoprosencephaly (a midline defect), we confirmed the breakpoints by performing FISH analysis using ZIC2 specific bacterial artificial chromosome (BAC) probes, RPCI 11 12-G12 selected from NCBI and Ensembl databases (BACPAC Resources, Children's Hospital Oakland Research Institute, Oakland, CA). This evidence concerns the gene ZIC2 and holoprosencephaly.