Figure 2. Detection of Arg348Cys mutation in the affected pedigree by direct sequencing and restriction fragment length polymorphism analysis

A: This panel shows the location of the 348th codon within SLC45A2. B: A representative chromatogram showing the location of the Arg348Cys mutation in heterozygous condition as indicated by the double peak (arrowhead). C: The allelic difference resulting from the Arg348Cys mutation was determined by BstU1 digestion of the PCR product obtained from exon 5 of the gene and the DNA digests of the five representative samples (four members of an affected family and a healthy control) were analyzed in a 6% polyacrylamide gel. The genotype of each individual is shown below the lane. The sizes of BstU1 uncut (236 bp) and digested (144 bp and 92 bp) DNA fragments are shown on the right side of the gel while the sizes (in bp) of the molecular weight marker (PBS X HaeIII) are shown on the left. Lanes 2 and 3 represent the DNA digests of two sibs affected with the mutation whereas lanes 1, 4, and 5 represent the father, mother. and healthy control, respectively.