Single locus microsatellites isolated using 5' anchored PCR.
Microsatellites are widely used as genetic markers because they are co-dominant, multiallelic, easily scored and highly polymorphic. A major drawback of microsatellite markers is the time and cost required to characterise them. We have developed a novel technique to reduce this cost by producing a microsatellite-rich PCR profile from genomic DNA which was cloned to yield a genomic library enriched for microsatellites. Sequence data and subsequent allele scoring within pedigrees revealed that these microsatellites retained their original repeat length and segregated normally. This technique permits genomic amplification with only one specific primer. Together with enrichment, the savings in primer costs reduces the cost of microsatellite characterisation considerably.
Fisher
P J
PJ
New Zealand Forest Research Institute Ltd, Rotorua. fisherp@tawa.fri.cri.nz
Gardner
R C
RC
Richardson
T E
TE
D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D018895:Microsatellite Repeats / Q000235:genetics*; D010944:Plants / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic
D017931:DNA Primers