Suppl Fig. 2

1. Supplemental Figure 2. mPCR-CE detects low abundance alleles, and can identify a 10% mass fraction of a fully unmethylated full mutation clinical sample in the background of a 90% mass fraction of an unmethylated full mutation clinical sample. Electropherograms and agarose gel images (outside panels) for A) sample #08, fully methylated, B) sample #125, unmethylated and C) a mixture of 10% of #08 in a background of 90% of #125. The inset data for sample #125 (Panel B) highlights the detection of CGG repeat peaks with a minor premutation allele at 160 CGG. This allele was detected with 60% methylation. In the mixed sample (Panel C), the recovery of 10% methylation of the full mutation allele was in agreement with the expected values. The agarose gel results, PCR products from undigested DNA (left ) and PCR products from HpaII digested DNA (right), were in complete agreement with the CE results (inset). These data confirm that mPCR-CE is fully capable of detecting low percent methylation in clinical samples with methylation mosaicism in the full mutation allele. The truncated “0-CGG” DNA” control amplicon is identified as “Ctrl” in each panel.

2. Suppl Fig. 2 Control - HEX HpaII Digested - FAM L AGE AGE L A) #08 Ctrl Ctrl B) #125 Ctrl Ctrl Normal to premutation C) #08 (10%) and #125 (90%) Ctrl Ctrl Ctrl Normal to premutation 10% Methylated