COMPLEX PROBES FOR HIGH-THROUGHPUT PARALLEL GENETIC-MAPPING OF GENOMIC MOUSE BAC CLONES

We describe a novel approach for the identification and mapping of pol ymorphic markers. Amplicons are generated by ligation of double-strand ed adaptor molecules to genomic DNA cleaved with a restriction enzyme. Using primers that extend beyond the restriction site, reduced-comple xity subsets of fragments are generated by PCR. Differences in the com position of complex probes generated from DNA of different strains are revealed through hybridization against high-density filter grids of l arge-insert genomic clones. Genetic mapping of genomic clones is achie ved by hybridizing complex probes derived from backcross animals again st the polymorphic clones. The mouse was chosen as a model system to t est the feasibility of this technique because of the general availabil ity of backcross resources and genomic libraries. Nevertheless, we wou ld expect the method to be of particular use to generate markers for s pecies that have not yet been extensively studied, because a substanti al number of easy-to-use markers can be recruited in a relatively shor t period of time.