title: Improved analysis of microsatellites using mass spectrometry.
abstract: The primer oligo base extension reaction combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, recently introduced by our group for detection of single-point mutations and small insertions/deletions, has been applied to the reliable quantification of nucleotide repeat units in microsatellites. The AluVpA DNA marker within intron 5 of the interferon-alpha receptor gene was chosen as the model system. By varying the dNTP/ddNTP mixtures used, the assay could also be directed to detect the location of second-site mutations within the repeats, resulting in identification of alleles not detectable by electrophoretic sizing methods and thus an increase of the polymorphism information content for a sampling of 28 unrelated individuals. The method results in highly informative mass signals and has the potential to increase the polymorphism information content for systems containing second-site mutations; thus it is a very attractive alternative technique in statistics-based gene mapping, cancer diagnostics, and forensic applications.
authors.affiliation: Sequenom, Inc., San Diego, California 92121, USA. andib@sequenom-san.com
authors.forename: A
authors.initials: A
authors.lastname: Braun
authors.forename: D P
authors.initials: DP
authors.lastname: Little
authors.forename: D
authors.initials: D
authors.lastname: Reuter
authors.forename: B
authors.initials: B
authors.lastname: Müller-Mysok
authors.forename: H
authors.initials: H
authors.lastname: Köster
mesh_terms: D001483:Base Sequence; D002648:Child; D004247:DNA / Q000032:analysis*; D017931:DNA Primers; D005260:Female; D005787:Gene Frequency; D005796:Genes / Q000235:genetics; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D007438:Introns / Q000235:genetics; D008297:Male; D018895:Microsatellite Repeats / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic; D053633:Receptor, Interferon alpha-beta; D017471:Receptors, Interferon / Q000235:genetics*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D017931:DNA Primers; D017471:Receptors, Interferon; D053633:Receptor, Interferon alpha-beta; D004247:DNA