Complex probes for high-throughput parallel genetic mapping of genomic mouse BAC clones.
We describe a novel approach for the identification and mapping of polymorphic markers. Amplicons are generated by ligation of double-stranded adaptor molecules to genomic DNA cleaved with a restriction enzyme. Using primers that extend beyond the restriction site, reduced-complexity subsets of fragments are generated by PCR. Differences in the composition of complex probes generated from DNA of different strains are revealed through hybridization against high-density filter grids of large-insert genomic clones. Genetic mapping of genomic clones is achieved by hybridizing complex probes derived from backcross animals against the polymorphic clones. The mouse was chosen as a model system to test the feasibility of this technique because of the general availability of backcross resources and genomic libraries. Nevertheless, we would expect the method to be of particular use to generate markers for species that have not yet been extensively studied, because a substantial number of easy-to-use markers can be recruited in a relatively short period of time.
Max-Planck-Institute for Molecular Genetics, Ihnestr. 73, D-14195 Berlin Dahlem, Germany.
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H
Himmelbauer
I
I
Dunkel
G W
GW
Otto
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C
Burgtorf
L C
LC
Schalkwyk
H
H
Lehrach
D000818:Animals; D002874:Chromosome Mapping / Q000379:methods*; D003001:Cloning, Molecular / Q000379:methods; D003433:Crosses, Genetic; D005819:Genetic Markers; D015698:Genomic Library; D051379:Mice; D008805:Mice, Inbred A; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D009115:Muridae / Q000235:genetics; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic
D005819:Genetic Markers