Abstract

We describe a flexible and general strategy for converting a wheat RFLP-based assay into a PCR-based sequence-tagged site (STS), and have applied it to derive markers for a powdery mildew resistance gene present in a wheat-rye translocation. The concept is based on deriving PCR primers that amplify all of the homoeoloci defined by a single-copy cDNA sequence, and separating the resulting mixture of homoeoamplicons via single-stranded conformation polymorphism (SSCP) gels, which are able to detect minor differences between related DNA sequences. After their separation, the individual homoeoamplicons were sequenced and these were used to define nucleotide polymorphisms that could be exploited to design locus-specific PCR primers. In one case, we were able to demonstrate that the assay was allele specific

Published in

Genome
2003, Volume: 46, number: 1, pages: 19-27
Publisher: NATL RESEARCH COUNCIL CANADA

SLU Authors

• Merker, Arnulf

  • Department of Crop Science, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Agricultural Science


Publication identifier

DOI: https://doi.org/10.1139/G02-101

Permanent link to this page (URI)

https://res.slu.se/id/publ/1701