title: Characterization of human AFLP systems apolipoprotein B, phenylalanine hydroxylase, and D1S80.
abstract: Methodology is presented for amplified fragment length polymorphism (AFLP) typing using a nonisotopic, PCR protocol. Human variable number tandem repeat (VNTR) loci used for identification in forensic and paternity testing were optimized for reaction and thermal-cycling parameters. Loci analyzed were the apolipoprotein B (APOB) 3' hypervariable region (HVR), phenylalanine hydroxylase 3' HVR (PAH), and D1S80. Coamplification of a monomorphic beta-globin fragment serves as an amplification control. Biotin is integrated into PCR amplicon through primer incorporation. AFLP products undergo agarose gel electrophoresis and Southern transfer to a nylon membrane. Amplicons were detected using a streptavidin-enzyme conjugate. Either colorimetric- or chemiluminescent-developed bands are genotyped using locus-specific allele ladders with known VNTR repeat numbers. Using this methodology, we have successfully typed > 500 individuals from three population groups for each locus during data basing and casework.
authors.affiliation: Roche Diagnostic Systems, Inc., Branchburg, New Jersey 08876.
authors.forename: D
authors.initials: D
authors.lastname: Latorra
authors.forename: C M
authors.initials: CM
authors.lastname: Stern
authors.forename: M S
authors.initials: MS
authors.lastname: Schanfield
mesh_terms: D000483:Alleles; D001055:Apolipoproteins B / Q000235:genetics*; D001483:Base Sequence; D006358:Hot Temperature; D006801:Humans; D018598:Minisatellite Repeats; D008969:Molecular Sequence Data; D010651:Phenylalanine Hydroxylase / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic
chemical_list: D001055:Apolipoproteins B; D010651:Phenylalanine Hydroxylase