Supplementary Figure 4: Junction PCR.
From: Biallelic mutations in human DCC cause developmental split-brain syndrome
(a) Digital droplet PCR using probes located within intron 1 of DCC (chr18: 49869550 (hg19) and chr18: 49871307 (hg19)) confirmed the homozygous deletion in both affected siblings (II:5 and II:7 from Family 1) and showed that their unaffected mother (I:2) was heterozygous for the deletion. (b) Schematic of the DCC gene with the predicted deletion near exon 1 (grey boxes). SNPs in blue indicate where copy number 2 was detected on the SNP array, and SNPs in green indicate where copy number 0 was detected on the SNP array, indicative of maximum and minimum boundaries of the predicted deletion, respectively. Nested PCR primers spanning the predicted deletion are indicated by F →, R ←. (c) Amplification with primers F5+R2 and F6+R2 yielded PCR products containing the deletion breakpoint in both the affected siblings (II:5 and II:7) and their mother (I:2), but not from an unrelated control individual. (d) Amplification with primers F2+R0 and F1+R0 yielded PCR products in the control sample, but non-specific bands in the affected individual II:7, confirming that the region can be amplified in the control sample.
