A new approach to extending the wheat marker pool by anchored PCR amplification of compound SSRs.
A study was undertaken to determine the utility in bread wheat of anchored PCR for the development of single locus SSR markers targeted at compound repeat motifs. In anchored PCR, microsatellite amplification is achieved using a single primer complementary to the flanking sequence, and one which anchors to the repeat junction of the compound SSR. The recovery rate of useable markers was found to be similar (43%) to that reported for conventionally generated SSRs. Thus, anchored PCR can be used to reduce the costs of marker development, since it requires that only half the number of primers be synthesised. Where fluorescence-based platforms are used, marker deployment costs are lower, since only the anchoring primers need to be labelled. In addition, anchored PCR improves the recovery of useful markers, as it allows assays to be generated from microsatellite clones with repeat sequences located close to their ends, a situation where conventional PCR amplification fails as two flanking primers cannot be designed. Strategies to permit the large-scale development of compound SSR markers amplified by anchored PCR are discussed.
Hayden
M J
MJ
Plant Breeding Institute, Cobbitty, University of Sydney, PMB 11, Camden, NSW, 2570 Australia. matthewh@camden.usyd.edu.au
Stephenson
P
P
Logojan
A M
AM
Khatkar
D
D
Rogers
C
C
Koebner
R M D
RM
Snape
J W
JW
Sharp
P J
PJ
D000483:Alleles; D002874:Chromosome Mapping; D017931:DNA Primers; D004586:Electrophoresis; D005453:Fluorescence; D018598:Minisatellite Repeats / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D016622:Silver Staining; D014908:Triticum / Q000235:genetics*
D017931:DNA Primers