Fig. 1

Schematic representation of allelic instability testing by real time PCR based SNP detection. a. Possible genotypes, from top to bottom: homozygous allele 1 and homozygous allele 2 (both non-informative), heterozygous normal and heterozygous imbalance (a 3:1 ratio of allele 1 vs. 2 and vice versa as a conceptual example). The probe corresponding to allele 1 is labeled with a 5’ VIC reporter, the allele 2 probe is labeled with a 5’ FAM reporter. Both probes contain a 3’ black hole quencher (BHQ). A = Adenine, T = Thymine, G = Guanine and C = Cytosine. b. The corresponding real time PCR component plots of above genotypes of representative SNP 8. X-axis: number of cycles, Y-axis: fluorescence. The arrows indicate the approximate delta Rn of the respective VIC or FAM reporters. c. Resulting scatterplot of SNP 8 of encountered genotypes. Dotted ovals indicate hypothetical homo- and heterozygous clusters. X-axis: delta Rn VIC, Y-axis: delta Rn FAM