Isolation of 48 genetic markers appropriate for high throughput genotyping of inbred rat strains by B1 repetitive sequence-representational difference analysis.
Mapping of genetic suppressors, modifiers, and quantitative trait loci (QTLs) requires genetic markers that can be efficiently and inexpensively genotyped for a large number of individuals. To isolate rat genetic markers suitable for this purpose, representational difference analysis (RDA) was performed with amplicons prepared by PCR with the B1 repetitive sequence used as the primer (B1-amplicons). In total, 48 polymorphic DNA fragments were isolated by five series of RDA, subtracting the B1-amplicons prepared from an ACI/N (ACI) rat from those prepared from BUF/Nac (BUF), and vice versa. All the polymorphic fragments detected "presence-or-absence" polymorphisms with B1-amplicons prepared from ACI, BUF, and their F2 progeny, and each fragment was linkage mapped. Dot-blotting amplicons onto filters at a high density and hybridization of the filters with these B1-RDA markers made it possible to genotype a large number of rats simultaneously for multiple loci. These B1-RDA markers were polymorphic between two given inbred strains of rat at frequencies between 30% and 70%. This is the first report on the isolation of B1-RDA markers among inbred strains of rats.
Ushijima
T
T
Carcinogenesis Division, National Cancer Center Research Institute, 1-1 Tsukiji 5-chome, Chuo-ku, Tokyo 104-0045, Japan.
Nomoto
T
T
Sugimura
T
T
Housman
D E
DE
Nagao
M
M
D000818:Animals; D002874:Chromosome Mapping; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D017931:DNA Primers; D005260:Female; D005784:Gene Amplification; D005819:Genetic Markers; D005838:Genotype; D008297:Male; D016133:Polymerase Chain Reaction; D051381:Rats; D011912:Rats, Inbred ACI; D011914:Rats, Inbred BN; D011915:Rats, Inbred BUF; D011916:Rats, Inbred F344; D020332:Rats, Inbred LEC; D011917:Rats, Inbred Lew; D011919:Rats, Inbred Strains; D011920:Rats, Inbred WF; D017207:Rats, Sprague-Dawley; D012091:Repetitive Sequences, Nucleic Acid
D017931:DNA Primers; D005819:Genetic Markers; D004247:DNA