title: Universal fluorescent labeling (UFL) method for automated microsatellite analysis.
abstract: We have devised a novel method for automated microsatellite analysis using "universal" fluorescent labeling. This system is based on polymerase chain reactions driven by sequence-specific primers and a reporter primer labeled with a fluorescent dye at its 5' end. The forward sequence-specific primer is designed with a tag region bearing no homology to any human genomic sequence. Complementary tag sequences act as templates for the 6-carboxyfluorescein-labeled reporter primer, and those products can be analyzed with an autosequencer. The results we achieved with this assay system were consistent with the results of conventional assays using radioisotope-labeled primers, and diagnosis required less time. Furthermore, the fluorescent-labeled reporter primer is "universal" in that it can be used with different sequence-specific primers designed to carry the appropriate tag sequence at their 5'-ends. Our observations suggest that the "universal" fluorescent labeling method is an efficient tool for analyzing sequence variations in human DNA.
authors.affiliation: Department of Molecular Biology, Institute of Gerontology, Nippon Medical School, 1-396 Kosugi-cho, Nakahara-ku, Kawasaki 211-8533, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Shimizu
authors.forename: Natsuki
authors.initials: N
authors.lastname: Kosaka
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimada
authors.forename: Takemitsu
authors.initials: T
authors.lastname: Nagahata
authors.forename: Hironori
authors.initials: H
authors.lastname: Iwasaki
authors.forename: Hisaki
authors.initials: H
authors.lastname: Nagai
authors.forename: Tadayoshi
authors.initials: T
authors.lastname: Shiba
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Emi
mesh_terms: D017931:DNA Primers; D005452:Fluoresceins; D018895:Microsatellite Repeats; D010761:Phosphorus Radioisotopes; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D017931:DNA Primers; D005452:Fluoresceins; D010761:Phosphorus Radioisotopes; C024098:6-carboxyfluorescein