title: Mutational analysis of the von Willebrand factor gene in type 1 von Willebrand disease using conformation sensitive gel electrophoresis: a comparison of fluorescent and manual techniques.
abstract: Two versions of conformation sensitive gel electrophoresis, fluorescent (F-CSGE) and manual (M-CSGE) techniques, were compared for mutation analysis of the von Willebrand factor gene. 56 PCRs were used to amplify all 52 exons of the gene in seven type 1 von Willebrand disease cases, plus a healthy control. One hundred and ninety-two samples were analyzed on each F-CSGE gel, compared with 40 on M-CSGE. 125 amplicons revealed bandshifts using F-CSGE, but only 101 by M-CSGE. Five mutations were detected by both techniques. F-CSGE detected 45 different polymorphisms whereas M-CSGE detected only 39. F-CSGE is high-throughput and more sensitive than M-CSGE.
authors.affiliation: Academic Unit of Hematology, School of Medicine and Biomedical Science, University of Sheffield, UK. hashemisoteh@gmail.com
authors.forename: Mohammad
authors.initials: M
authors.lastname: Hashemi Soteh
authors.forename: Ian R
authors.initials: IR
authors.lastname: Peake
authors.forename: Luke
authors.initials: L
authors.lastname: Marsden
authors.forename: John
authors.initials: J
authors.lastname: Anson
authors.forename: Javier
authors.initials: J
authors.lastname: Batlle
authors.forename: Dominique
authors.initials: D
authors.lastname: Meyer
authors.forename: Edith
authors.initials: E
authors.lastname: Fressinaud
authors.forename: Claudine
authors.initials: C
authors.lastname: Mazurier
authors.forename: Jenny
authors.initials: J
authors.lastname: Goudemand
authors.forename: Jeroen
authors.initials: J
authors.lastname: Eikenboom
authors.forename: Anne
authors.initials: A
authors.lastname: Goodeve
mesh_terms: D004247:DNA / Q000097:blood / Q000737:chemistry / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005091:Exons / Q000235:genetics; D005456:Fluorescent Dyes / Q000032:analysis; D005470:Fluorometry; D020180:Heteroduplex Analysis / Q000379:methods*; D006801:Humans; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D011544:Pseudogenes; D012680:Sensitivity and Specificity; D014842:von Willebrand Diseases / Q000235:genetics*; D014841:von Willebrand Factor / Q000737:chemistry / Q000235:genetics*
chemical_list: D005456:Fluorescent Dyes; D014841:von Willebrand Factor; D004247:DNA