Detection of Large Pathogenic Expansions in FRDA1, SCA10, and SCA12 Genes Using a Simple Fluorescent Repeat-Primed PCR Assay Claudia Cagnoli, Chiara.

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Detection of Large Pathogenic Expansions in FRDA1, SCA10, and SCA12 Genes Using a Simple Fluorescent Repeat-Primed PCR Assay Claudia Cagnoli, Chiara Michielotto, Tohru Matsuura, Tetsuo Ashizawa, Russell L. Margolis, Susan E. Holmes, Cinzia Gellera, Nicola Migone, Alfredo Brusco The Journal of Molecular Diagnostics Volume 6, Issue 2, Pages 96-100 (May 2004) DOI: 10.1016/S1525-1578(10)60496-5 Copyright © 2004 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Fluorescent repeat-primed PCR analysis of the Friedreich ataxia locus. The peak profiles of healthy controls with two alleles in the normal size range (panels 1 and 2) are distinct from the profiles found in subjects with one (Hz carrier, panel 3) or two pathogenic alleles (FA patients, panels 4 and 5). The genotype of FRDA1 GAA triplets is shown inside each panel. Marker peaks of 250, 300, 340, and 350 bp are shadowed. Size in bp is also reported on abscissa, while ordinate shows an arbitrary fluorescence intensity scale. The hyphened arrow marks the right tail shape of the peak traces. The Journal of Molecular Diagnostics 2004 6, 96-100DOI: (10.1016/S1525-1578(10)60496-5) Copyright © 2004 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Measurement of the slope of the peaks profile. Height (F.U.) and size (bp) of repeat-primed PCR peaks can be plotted to calculated a regression line and an R2 coefficient (see Materials and Methods). A control subject and a patient corresponding to panels 2 and 5 in Figure 1 are displayed as example. The R2 values were always able to discriminate a carrier of one or two expansions versus a normal control (see Table 2). The Journal of Molecular Diagnostics 2004 6, 96-100DOI: (10.1016/S1525-1578(10)60496-5) Copyright © 2004 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Fluorescent repeat-primed PCR analysis in the SCA10 (left) and SCA12 (right) loci. Top panels show healthy control subjects, while bottom panels display patients carrying one expanded allele. The genotype at the two loci expressed in number of repeats is reported inside each panel. Marker peaks of 139, 150, 160, 200, 250, 300, 340, and 350 bp are shadowed. Size in bp is also reported on abscissa, while ordinate shows an arbitrary fluorescence intensity scale. The Journal of Molecular Diagnostics 2004 6, 96-100DOI: (10.1016/S1525-1578(10)60496-5) Copyright © 2004 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions