Figure 1 | Modern Pathology

Figure 1

From: Identification of SYT-SSX transcripts from synovial sarcomas using RT-multiplex PCR and capillary electrophoresis

Figure 1

Schematic diagram of RT-multiplex PCR for detection and genotypic characterization of SYT-SSX transcripts, depicting relative locations of primers, PCR amplicon sizes, and expected capillary electrophoresis results from tumors harboring either SYT-SSX1 or SYT-SSX2 transcripts. RNA extracted from FFPE tissue blocks is combined with all RT-PCR reagents in a single tube. A 176 bp NED-labeled amplicon is produced from GAPD transcripts and 6-FAM-labeled amplicons are produced from the SYT-SSX fusion transcripts, one (102 bp) common to SYT fused to any SSX gene, and a second unique to either SYT-SSX1 (146 bp) or SYT-SSX2 (130 bp) transcripts. Products are analyzed by capillary electrophoresis and genotype is assigned based on amplicon sizes. Relative primer locations are depicted by chevrons, C indicates a common priming site, 1 and 2 are unique sites. Locations of nucleotides, which differ between exon 5 of SSX1 and SSX2 are shown by vertical lines. Labels on the 5′ ends of primers and amplicons are represented as black (6-FAM) or gray (NED).

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