title: A simple method to confirm and size deletion, duplication, and insertion mutations detected by sequence analysis.
abstract: Characterizing heterozygous insertions or deletions in genes by PCR and Sanger sequencing can be a challenge due to overlapping sequencing traces produced by overlapping templates. This is particularly problematic for clinical diagnostic laboratories, because mutations must be precisely characterized. Although the mutation detection software used by clinical diagnostic laboratories reliably identifies small insertions and deletions, overlapping deletions and insertions on opposite chromosomes, complex rearrangements, and insertions or deletions close to the primer sites may be missed. Here we describe a rapid, simple method to confirm and precisely characterize deletions and insertions using a capillary-based gel electrophoresis system. This technique has been applied to a series of patients with deletion, duplication, or insertion mutations identified by sequencing, as well as to patients with repeat tract polymorphisms, to demonstrate the utility of this method.
authors.affiliation: Department of Human Genetics, Emory University, Atlanta, Georgia 30322, USA.
authors.forename: Lawrence N
authors.initials: LN
authors.lastname: Hjelm
authors.forename: Ephrem L H
authors.initials: EL
authors.lastname: Chin
authors.forename: Madhuri R
authors.initials: MR
authors.lastname: Hegde
authors.forename: Bradford W
authors.initials: BW
authors.lastname: Coffee
authors.forename: Lora J H
authors.initials: LJ
authors.lastname: Bean
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D003638:Deafness / Q000235:genetics; D006801:Humans; D009154:Mutation; D009542:Niemann-Pick Diseases / Q000235:genetics; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers