Analysis of the relative abundance of wild-type and mutant mRNAs.

(A) Example of GeneScan analysis window showing fluorescence peaks corresponding to wild-type and mutant fragments for Arg110stop mutation. These fragments, whose lengths are indicated below each peak, were obtained by a suitable allele-specific digestion subsequent to an RT-PCR performed on RNA extracted from transfected COS-1 cells, coexpressing wild type and each mutant Aα-chain (see “Materials and methods”). The x-axis represents GeneScan data points, and the y-axis represents FUs. (B) Comparison of wild-type and mutant mRNAs levels. The reported values correspond to the peak area measured by means of GeneScan analysis software, with the wild-type mRNA level set equal to 100%.