title: Towards fully automated genotyping: use of an X linked recessive spastic paraplegia family to test alternative analysis methods.
abstract: Advances in dinucleotide-based genetic maps open possibilities for large scale genotyping at high resolution. The current rate-limiting steps in use of these dense maps is data interpretation (allele definition), data entry, and statistical calculations. We have recently reported automated allele identification methods. Here we show that a 10-cM framework map of the human X chromosome can be analyzed on two lanes of an automated sequencer per individual (10-12 loci per lane). We use this map and analysis strategy to generate allele data for an X-linked recessive spastic paraplegia family with a known PLP mutation. We analyzed 198 genotypes in a single gel and used the data to test three methods of data analysis: manual meiotic breakpoint mapping, automated concordance analysis, and whole chromosome multipoint linkage analysis. All methods pinpointed the correct location of the gene. We propose that multipoint exclusion mapping may permit valid inflation of LOD scores using the equation max LOD-(next best LOD).
authors.affiliation: Department of Molecular Genetics and Biochemistry, Human Genetics and Pediatrics, University of Pittsburgh School of Medicine, PA 15261, USA.
authors.forename: H
authors.initials: H
authors.lastname: Kobayashi
authors.forename: T C
authors.initials: TC
authors.lastname: Matise
authors.forename: M W
authors.initials: MW
authors.lastname: Perlin
authors.forename: H G
authors.initials: HG
authors.lastname: Marks
authors.forename: E P
authors.initials: EP
authors.lastname: Hoffman
mesh_terms: D000483:Alleles; D001331:Automation; D002874:Chromosome Mapping / Q000379:methods*; D004247:DNA / Q000032:analysis; D005260:Female; D005808:Genes, Recessive; D006580:Genetic Carrier Screening; D008040:Genetic Linkage / Q000235:genetics*; D005838:Genotype; D006801:Humans; D008126:Lod Score; D008297:Male; D008540:Meiosis; D010375:Pedigree; D016133:Polymerase Chain Reaction; D015419:Spastic Paraplegia, Hereditary / Q000235:genetics*; D014960:X Chromosome
chemical_list: D004247:DNA