Genomic DNA (gDNA) and complementary DNA (cDNA) sequencing of mesenchymal stem cells samples from four TCS patients. The pathogenic mutation of TCS patients 16, 21, 22, and 23 are c.218_222insAACC (exon 3), c.4344dupA (exon 24), c.431delC (exon 5), and c.4218dupG (exon 23), respectively. gDNA was sequenced with intronic primers and cDNA with exonic primers. Observing all gDNA samples, we can assume that all analyzed individuals are heterozygous for the pathogenic mutation. Analyzing cDNA samples, we could detect the mutant allele expression in TCS 16 and 23; TCS 21 and 22 express only the wild-type allele. Note that even when the mutant allele is expressed (patients TCS16 and TCS23), the peak heights are lower.
Masotti et al. BMC Medical Genetics 2009 10:136 doi:10.1186/1471-2350-10-136