Automated single‐strand conformation polymorphism reveals low diversity of a Major Histocompatibility Complex Class II gene in the threatened New Zealand sea lion |
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Authors: | G. M. Lento C. S. Baker V. David N. Yuhki N. J. Gales S. J. O'Brien |
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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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Keywords: | ABI DQB genetic diversity Major Histocompatibility Complex New Zealand sea lion population genetics SSCP |
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