167

1. I II bp eIVS5-16G>A(MspI cut) 201 167 103 34 e911delT(PmlI cut) bp 705/704 427 277 Supplemental figure (E)F-1: Restriction enzyme analysis. To detect the CHRNE mutation IVS5-16G>A, a 304 bp PCR fragment containing the mutation in intron 5 was amplified and digested with MspI. The digest yields 167, 103, and 34 bp fragments for the wild-type allele. As the mutation IVS5-16G>A abolishes a MspI restriction site, digest of the mutated allele results in fragments of 201 and 103 bp length. The patient (II:2), his healthy sister (II:1) and the mother (I:1) carry the mutation heterozygously whereas the father (I:2) does not carry the mutation. For the mutation 911delT a 705/704 bp fragment was amplified. The mutation 911delT creates a new PmlI site. The 705 bp wild-type allele remains undigested, whereas the 704 bp mutant allele yields two fragments (277 bp and 427 bp). The father (I:2) and the patient (II:2) carry the mutation heterozygously. Only the patient harbors both mutated alleles.