Figure 4 | Scientific Reports

Figure 4

From: Development and validation of a 4-color multiplexing spinal muscular atrophy (SMA) genotyping assay on a novel integrated digital PCR instrument

Figure 4

Representative partition image (A) and scatter plots (B–D) of the SMA 4-color multiplexing dPCR assay using NA03815 genomic DNA as the sample. The scatter plots are shown for SMN1 exon 7 (B; FAM), SMN2 exon 7 (C; VIC) and total SMN intron 1 (D; TYE665), all relative to the reference gene RPPH1 (TAMRA). The analysis software subtracted the pre-PCR image intensities from the post-PCR image intensities so as to remove fluorescent signals which did not exhibit amplification behavior, thereby eliminating false positive signals. If the pre-PCR partitions were brighter than the post-PCR partitions, then the analysis software would record negative fluorescent units. These negative units, however, did not impact the copy number quantification results.

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