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. 2010 Aug 24;5(8):e12315. doi: 10.1371/journal.pone.0012315

Figure 5. Triplex competitive RT-PCR assay to evaluate trace levels of Math5 splicing in the embryonic retina.

Figure 5

A. Diagram showing PCR strategy. The length and %G+C of competing amplicons are comparable. B. Agarose gel stained with ethidium bromide, showing only the unspliced Math5 cDNA product in each assay. C. Capillary electrophoresis profiles showing triplex competitive RT-PCR products (top panels) and the ECO product amplified with duplex UTR primers in the presence of 1X MA (bottom panel). The common antisense primer (LP4) was end-labeled with 6-FAM. From the peak areas measured in replicate experiments and mixing controls, we estimate that the ECO product represents 0.4 to 1.0 percent of Math5 mRNA in the embryonic retina, which is near the detection limit of this assay.