Sequence tagged microsatellite profiling (STMP): improved isolation of DNA sequence flanking target SSRs.
Sequence tagged microsatellite profiling (STMP) enables the rapid development of large numbers of co-dominant DNA markers, known as sequence tagged microsatellites (STMs). Each STM is amplified by PCR using a single primer specific to the conserved DNA sequence flanking the microsatellite repeat in combination with a universal primer that anchors to the 5'-ends of the microsatellites. It is also possible to convert STMs into conventional microsatellite, or simple sequence repeat (SSR), markers that are amplified using a pair of primers flanking the repeat sequence. Here, we describe a modification of the STMP procedure to significantly improve the capacity to convert STMs into conventional SSRs and, therefore, facilitate the development of highly specific DNA markers for purposes such as marker-assisted breeding. The usefulness of this technique was demonstrated in bread wheat.
Hayden
M J
MJ
Plant Breeding Institute, University of Sydney, PMB 11, Camden, NSW 2570, Australia. matthewh@camden.usyd.edu.au
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D017124:Conserved Sequence; D004247:DNA / Q000302:isolation & purification*; D017931:DNA Primers / Q000737:chemistry; D004262:DNA Restriction Enzymes; D005819:Genetic Markers; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction / Q000379:methods*; D016324:Sequence Tagged Sites
D017931:DNA Primers; D005819:Genetic Markers; D004247:DNA; D004262:DNA Restriction Enzymes