Journal
MOLECULAR ECOLOGY NOTES
Volume 3, Issue 3, Pages 346-349Publisher
BLACKWELL PUBLISHING LTD
DOI: 10.1046/j.1471-8286.2003.00445.x
Keywords
ABI; DQB; genetic diversity; major histocompatibility complex; new Zealand sea lion; population genetics; SSCP
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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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