Fig. 2

ARMS‐qPCR for the point mutation c.4789C > T (p.Arg1597Trp) of VWF gene. A standard curve was generated by plotting the difference between the PCR cycle crossing point (Cp) of the wild-type (WT) allele and the mutant (MU) allele i.e. ΔCp (WT-MU) value of different synthetic dilutions against the mutant % of the synthetic dilution. The synthetic dilution were prepared by a 2-fold serial dilution of MU DNA by WT DNA. (a). According to the S‐curve formula, amplification plot was analyzed at genomic level of an affected family with the paternally‐inherited c.4789C > T mutation to detect the mutant % in different samples of maternal blood (b), the proband (affected children) blood (c), paternal blood (d) and oral mucosal cells (e), and the results showed 0 %, 49 %, 25.5 % and 31.1 % in mutant percentage, respectively. Triplicated ARMS‐qPCR tests with WT allele‐specific primers are indicated by WT lines in one color, whereas the tests with MU allele‐specific primers are indicated by MU lines in another color