Figure 1.

Workflow for amplification and detection of FMR1 amplicons using a three-primer FMR1 PCR. Input gDNA is amplified by two gene-specific primers (forward [Fwd] and reverse [Rev]) and a CGG repeat primer in a single tube. After amplification, the products, which include the full-length amplicon that completely encompasses the triplet repeat region and a multiplicity of CGG repeat primed products, are resolved by CE. The resulting electropherogram supports quantification of the number of CGG repeats, determination of the allele zygosity, and the sequence context of any AGG spacer elements.