A sensitive method for detecting variation in copy numbers of duplicated genes. Academic Article uri icon


  • Gene duplications are common in the vertebrate genome, and duplicated loci often show a variation in copy number that may have important phenotypic effects. Here we describe a powerful method for quantification of duplicated copies based on pyrosequencing. A reliable quantification was obtained by amplification of the duplication break-point and a corresponding nonduplicated sequence in a competitive PCR assay. A comparison with an independent method for quantification based on the Invader technology revealed an excellent correlation between the two methods. The pyrosequencing-based method was evaluated by analyzing variation in copy number at the duplicated KIT/Dominant white locus in pigs. We were able to distinguish haplotypes at this locus by combining the duplication breakpoint test with a diagnostic test for a functionally important splice mutation in the duplicated gene. An extensive allelic variation, including the presence of a new allele carrying a single KIT copy expected to encode a truncated KIT receptor, was revealed when analyzing white pigs from commercial lines.

published proceedings

  • Genome Res

altmetric score

  • 3

author list (cited authors)

  • Pielberg, G., Day, A. E., Plastow, G. S., & Andersson, L.

citation count

  • 44

complete list of authors

  • Pielberg, Gerli||Day, Andy E||Plastow, Graham S||Andersson, Leif

publication date

  • September 2003