PCR digestion analysis in family A.

DNA was prepared from peripheral blood for patients I 1 and I 2. Patient II 1 died in the perinatal period and was not available for the study. For the fetus (II 2), DNA was obtained from CVS. “nal” indicates normal control sample, MWS (molecular weight standard) = ΦX174 RF/HaeIII fragments. (A) Pedigree of family A. (B) PCR digestion of exon 3. Amplification of exons 2, 3, and 4 with the 1838 F and 2549 R set of primers was followed by digestion with DdeI (normal fragment 206 bp, mutants fragments 178 and 28 bp). Patients I 1 and II 2 are heterozygous for the mutation; patient I 2 is normal. (B) PCR digestion of exon 1. Amplification with the 520 F and 776 R set of primers followed by digestion with MwoI (normal fragment 190 bp, mutant fragments 133 and 56 bp). Patients I 1 and II 2 are normal, and patient I 2 is heterozygous for the mutation del 86-87.