Suppl Fig. 2

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.

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Suppl Fig. 2

Normal to premutation10% Methylated

Control - HEX HpaII Digested - FAML AGE AGE L

A) #08

B) #125

C) #08 (10%) and #125 (90%)

Normal to premutation

Ctrl

CtrlCtrl

Ctrl

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