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    Multiplex PCR and minisequencing of SNPs--a model with 35 Y chromosome SNPs

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    Reference Type: Journal Article

    Record Number: 136

    Author: J. J. Sanchez, C. Borsting, C. Hallenberg, A. Buchard, A. Hernandez and N. Morling

    Year: 2003

    Title: Multiplex PCR and minisequencing of SNPs--a model with 35 Y chromosome SNPs

    Journal: Forensic Sci Int

    Volume: 137

    Issue: 1

    Pages: 74-84

    Epub Date: 2003/10/11

    Date: Oct 14

    Short Title: Multiplex PCR and minisequencing of SNPs--a model with 35 Y chromosome SNPs

    ISSN: 0379-0738 (Print)

    DOI: S0379073803002998 [pii],

    Accession Number: 14550618

    Keywords: *Chromosomes, Human, Y, DNA/analysis, DNA Primers, Denmark, Electrophoresis, Capillary, Female, Fluorescence, Humans, Male, Models, Genetic, Polymerase Chain Reaction/*methods, *Polymorphism, Single Nucleotide, Sequence Analysis, DNA/*methods,

    Abstract: We have developed a robust single nucleotide polymorphism (SNPs) typing assay with co-amplification of 25 DNA-fragments and the detection of 35 human Y chromosome SNPs. The sizes of the PCR products ranged from 79 to 186 base pairs. PCR primers were designed to have a theoretical Tm of 60 +/- 5 degrees C at a salt concentration of 180 mM. The sizes of the primers ranged from 19 to 34 nucleotides. The concentration of amplification primers was adjusted to obtain balanced amounts of PCR products in 8mM MgCl2. For routine purposes, 1 ng of genomic DNA was amplified and the lower limit was approximately 100 pg DNA. The minisequencing reactions were performed simultaneously for all 35 SNPs with fluorescently labelled dideoxynucleotides. The size of the minisequencing primers ranged from 19 to 106 nucleotides. The minisequencing reactions were analysed by capillary electrophoresis and multicolour fluorescence detection. Female DNA did not influence the results of Y chromosome SNP typing when added in concentrations more than 300 times the concentrations of male DNA. The frequencies of the 35 SNPs were determined in 194 male Danes. The gene diversity of the SNPs ranged from 0.01 to 0.5.

    Notes: Sanchez, Juan J, Borsting, Claus, Hallenberg, Charlotte, Buchard, Anders, Hernandez, Alexis, Morling, Niels, Research Support, Non-U.S. Gov't, Ireland, Forensic science international, Forensic Sci Int. 2003 Oct 14;137(1):74-84.,

    Author Address: Department of Forensic Genetics, Institute of Forensic Medicine, University of Copenhagen, 11 Frederik V's Vej, DK-2100 Copenhagen, Denmark. juan.sanchez@forensic.ku.dk

    Language: eng