Fig. 3
From: Detection of Novel LAMC2 Mutations in Herlitz Junctional Epidermolysis Bullosa
Demonstration of the maternal LAMC2 mutation in Family 1
(A) Conformation sensitive gel electrophoresis (CSGE) of exon 19 and flanking intronic sequences reveals broadened bands with the mother’s (M) and the proband’s (P) DNA, when compared with DNA from the father (F) and normal controls (not shown). (B) Direct nucleotide sequencing of the proband’s PCR product reveals a heterozygous G-to-A transition at the 5′ donor splice site of intron 19 (upper panel) when compared with the normal sequence (lower panel). Upper case represents the coding sequence; lower case represents the intronic sequences. (C) The mutation was verified by allele-specific oligonucleotide hybridization (ASO) with oligomers corresponding to the wild-type (WT) and mutant (M) alleles and using PCR products of the father’s (F), mother’s (M), proband’s (P), and normal control (C) DNA. The sequences for oligomers used for hybridization were: WT, 5′-ACCTCAGAG-GTTAGTACTTC-3′; M, 5′-ACCTCAGAGATTAGTACTTC-3′. The Tm values of these oligomers were 58°C and 56°C, respectively.