Construction of a high-throughput rat genetic mapping system with 466 arbitrarily primed-representational difference analysis markers.
Linkage mapping of quantitative trait loci (QTLs) requires genetic markers that can be efficiently genotyped for a large number of individuals. To isolate genetic markers suitable for this purpose, we previously established the arbitrarily primed RDA (AP-RDA) method. Dot-blotting AP-PCR products (AP-amplicons) onto filters at a high density and hybridization of the filters with the AP-RDA markers made it possible to genotype a large number of individuals simultaneously for multiple loci. In this study, by using 25 primers or primer combinations, we isolated a total of 419 AP-RDA markers by subtracting the AP-amplicon of BUF rats from that of ACI rats, and vice versa. By combining 47 previously isolated markers, a rat genetic map was drawn with 466 AP-RDA markers. Between two given strains of rats other than ACI and BUF, the average informativeness of the markers was 38%. As for the intercross of ACI and BUF rats, 12 selected primers served to genotype 259 loci. In addition, the amounts and quality of genomic DNA to be used for AP-PCR were examined to guarantee reliable genotyping. Now, initial genome scanning of the rat for linkage analysis can be performed efficiently using this mapping system with AP-RDA markers.
Yamashita
S
S
Carcinogenesis Division, National Cancer Center Research Division, Tokyo, Japan.
Yoshida
Y
Y
Kurahashi
A
A
Sugimura
T
T
Ushijima
T
T
D000818:Animals; D002874:Chromosome Mapping / Q000379:methods*; D004247:DNA / Q000032:analysis / Q000235:genetics; D008040:Genetic Linkage; D005819:Genetic Markers; D051381:Rats; D011919:Rats, Inbred Strains / Q000235:genetics*; D017208:Rats, Wistar
D005819:Genetic Markers; D004247:DNA