Characterization of human AFLP systems apolipoprotein B, phenylalanine hydroxylase, and D1S80.
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.
Latorra
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Roche Diagnostic Systems, Inc., Branchburg, New Jersey 08876.
Stern
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Schanfield
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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
D001055:Apolipoproteins B; D010651:Phenylalanine Hydroxylase