title: Direct detection and automated sequencing of individual alleles after electrophoretic strand separation: identification of a common nonsense mutation in exon 9 of the human lipoprotein lipase gene.
abstract: Large-scale screening by direct sequencing of DNA to detect molecular variants remains a laborious endeavor whose difficulty is compounded by heterozygosity. We show that mobility shifts of single-stranded DNA electrophoresed under nondenaturing conditions can be used not only to detect variants (Orita,M. et al., 1989, Genomics, 5, 874-879), but also to separate and sequence directly individual alleles. In this manner, we have identified a common variant of human lipoprotein lipase resulting from a nonsense mutation in exon 9 of the gene. Whether this variant is of functional significance remains to be determined.
authors.affiliation: Howard Hughes Medical Institute, University of Utah Health Sciences Center, Salt Lake City 84132.
authors.forename: A
authors.initials: A
authors.lastname: Hata
authors.forename: M
authors.initials: M
authors.lastname: Robertson
authors.forename: M
authors.initials: M
authors.lastname: Emi
authors.forename: J M
authors.initials: JM
authors.lastname: Lalouel
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D004586:Electrophoresis; D005091:Exons; D005796:Genes; D006579:Heterozygote; D006801:Humans; D015228:Hypertriglyceridemia / Q000201:enzymology / Q000235:genetics; D008071:Lipoprotein Lipase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction
chemical_list: D008071:Lipoprotein Lipase