Figure 2

Allele-specific amplification ofTSC2mosaic variants. a. Allele-specific amplification of the TSC2 c.3099C>G (p.Y1033*) mosaic variant. Specific amplification of the mutant c.3099G (upper panel) and normal c.3099C (lower panel) alleles from DNA from an unrelated healthy individual (control), DNA from individual III (III), both parents, and an individual with TSC heterozygous for the TSC2 c.3099C>G (p.Y1033*) pathogenic variant (c.3099C/G). b. Allele-specific amplification of the TSC2 c.2838-122G>A mosaic variant. Specific amplification of the mutant c.2838-122A (upper panel) and normal c.2838-122G (lower panel) alleles from DNA from an unrelated healthy individual (control), individual VI (VI) and both parents. c. RT-PCR of TSC2 mRNA from individual VI (VI) and 5 unrelated individuals (controls). An extra splice variant was amplified from RNA from individual VI (arrow), but not from the controls. d. Sequence of the additional RT-PCR product identified in individual VI (see Figure 2c, arrow). Sequences derived from exons 25 and 26 are in capitals; the premature stop codon is underlined. e. PCR amplification of the TSC2 c.226-1086del10 (intron 3; chr16 g.2102256del10) variant (rs140492671) from DNA from individuals IV and VII, the parents of individual IV and DNA from an unrelated healthy individual (control).