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Article
Automated Single-Strand Conformation Polymorphism Reveals Low Diversity of a Major Histocompatibility Complex Class II Gene in the Threatened New Zealand Sea Lion
Molecular Ecology Resources
  • G. M. Lento, University of Auckland
  • C. S. Baker, University of Auckland
  • Victor David, National Cancer Institute at Frederick
  • Naoya Yuhki, National Cancer Institute at Frederick
  • N. J. Gales, Australian Antarctic Division
  • Stephen J. O'Brien, National Cancer Institute at Frederick
Document Type
Article
Publication Date
9-1-2003
Keywords
  • ABI,
  • DQB,
  • Genetic diversity,
  • Major Histocompatibility Complex,
  • New Zealand sea lion,
  • Population genetics,
  • SSCP
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.

Comments

©2003 Blackwell Publishing Ltd

ORCID ID
0000-0001-7353-8301
ResearcherID
N-1726-2015
Citation Information
G. M. Lento, C. S. Baker, Victor David, Naoya Yuhki, et al.. "Automated Single-Strand Conformation Polymorphism Reveals Low Diversity of a Major Histocompatibility Complex Class II Gene in the Threatened New Zealand Sea Lion" Molecular Ecology Resources Vol. 3 Iss. 2 (2003) p. 346 - 349 ISSN: 1755-098X
Available at: http://works.bepress.com/stephen-obrien/84/