title: High-resolution melting analysis for genotyping of the myotonic dystrophy type 1 associated Alu insertion/deletion polymorphism.
abstract: Since its introduction, high-resolution melting (HRM) analysis has been used for genotyping of various types of sequence alterations. In this study, we report the use of HRM for genotyping of the 1-kb insertion/deletion polymorphism, involving a problematic region of five consecutive Alu elements, that is associated with myotonic dystrophy type 1. We combined a three-primer polymerase chain reaction (PCR) amplification approach with HRM using two primer sets. Analyses based on curve shapes are sensitive enough to differentiate between genotypes with both primer sets. In addition, the newly designed insertion-specific primer from the second primer set equalizes the allele-specific amplicon lengths, thereby reducing the possibility of preferential amplification of shorter fragments.
authors.affiliation: Department of Molecular Biology, Faculty of Natural Sciences, Comenius University, 842 15 Bratislava 4, Slovakia. jradvanszky@gmail.com
authors.forename: Jan
authors.initials: J
authors.lastname: Radvansky
authors.forename: Peter
authors.initials: P
authors.lastname: Resko
authors.forename: Milan
authors.initials: M
authors.lastname: Surovy
authors.forename: Gabriel
authors.initials: G
authors.lastname: Minarik
authors.forename: Andrej
authors.initials: A
authors.lastname: Ficek
authors.forename: Ludevit
authors.initials: L
authors.lastname: Kadasi
mesh_terms: D020087:Alu Elements / Q000235:genetics*; D017353:Gene Deletion; D005838:Genotype; D006801:Humans; D009223:Myotonic Dystrophy / Q000175:diagnosis / Q000235:genetics*; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D044366:Transition Temperature