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.
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.
Hashemi Soteh
Mohammad
M
Academic Unit of Hematology, School of Medicine and Biomedical Science, University of Sheffield, UK. hashemisoteh@gmail.com
Peake
Ian R
IR
Marsden
Luke
L
Anson
John
J
Batlle
Javier
J
Meyer
Dominique
D
Fressinaud
Edith
E
Mazurier
Claudine
C
Goudemand
Jenny
J
Eikenboom
Jeroen
J
Goodeve
Anne
A
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*
D005456:Fluorescent Dyes; D014841:von Willebrand Factor; D004247:DNA