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Automated single-strand conformation polymorphism reveals low diversity of a Major Histocompatibility Complex Class II gene in the threatened New Zealand sea lion

  1. Author:
    Lento, G. M.
    Baker, C. S.
    David, V.
    Yuhki, N.
    Gales, N. J.
    O'Brien, S. J.

  2. Author Address

    Univ Auckland, Sch Biol Sci, Populat Genet & Evolut Res Grp, Private Bag 92019, Auckland 1, New Zealand Univ Auckland, Sch Biol Sci, Populat Genet & Evolut Res Grp, Auckland 1, New Zealand NCI, Frederick Canc Res & Dev Ctr, Div Basic Sci, Lab Genom Divers, Frederick, MD 21702 USA Australian Antarctic Div, Kingston, Tas 7050, Australia Baker CS Univ Auckland, Sch Biol Sci, Populat Genet & Evolut Res Grp, Private Bag 92019, Auckland 1, New Zealand
    1. Year: 2003
  2. Journal: Molecular Ecology Notes
    1. 3
    2. 3
    3. Pages: 346-349
  4. Type of Article: Article
  1. Abstract:

    Single-strand conformation polymorphism (SSCP) analysis of the second exon of the DQB gene in the endemic New Zealand sea lion (Phocarctos hookeri) was automated using fluorescently-labelled PCR primers and a capillary sequencer. SSCP mobility profiles (n = 61 individuals) were confirmed by direct sequencing of genomic polymerase chain reaction (PCR) products (n = 39). We found only two alleles defined by changes at a single codon of this apparently functional locus. SSCP at a constant temperature above ambient allowed consistent and rapid genotyping. By comparison, sequence-based genotyping was highly dependent on the threshold used to identify secondary peaks indicative of a heterozygote.

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