Fig. 1

Structural variation detected in WHO reference samples. a, b, c, and d Paralog-specific analyses (top) with corresponding quantitative multiplex PCR of short fluorescent fragments (QMPSF) results (below) for RBC1, RBC5, RBC4, and RBC12, respectively. Each paralog-specific panel shows scale RHD (blue) and RHCE (red) gene schematics (top) and the location of single unique nucleotides within genic regions (black) and in Rhesus boxes (gray). Gray circles within panels represent normalized mean read depth for k-mers corresponding to singly unique nucleotides (SUNs). The dashed gray line denotes a copy number of 2; solid blue and red lines indicate inferred copy numbers over the RHD and RHCE genes, respectively. In QMPSF panels, peak heights are fluorescence measurements normalized to the amplified exon for HFE. The F9 peak serves as an additional positive amplification control. Light yellow panels in QMPSF results for b RBC5 and d RBC12 highlight RHD whole-gene deletions.  Red asterisks highlight amplicons with results indicative of structural variation. Note: In d, QMPSF amplification of exon 8 is suggestive of structural variation, although exon 8 by next-generation sequencing (NGS) analyses is predicted to be unaffected