Analysis of a set of RAPD markers by hybridization and sequencing in Brassica: a note of caution.

A series of RAPD markers generated by a single 10-mer primer were analyzed by hybridization to amplified and genomic DNA and by sequencing in two Brassica species. Primer B18 produced different profiles of nine major bands each in both Brassica nigra (B genome) and B. napus (AC genomes). Cloning and sequencing of five B18 B. nigra amplification products revealed that they were all unrelated to each other. Only limited stretches of high similarity of up to 69 nucleotides were shared by some of these clones. Hybridization to genomic DNA indicated that only two corresponded to a highly repeated sequence, whereas the rest were low copy sequences. In spite of their lack of homology, when these clones were used as probes to amplified B. nigra DNA, they hybridized to multiple bands in the profile. Hybridization of B. nigra clones for bands of similar sizes in both species, failed to hybridize in B. napus, revealing lack of homology between the DNAs of the two species. Because of these inconsistencies, it is concluded that RAPD markers, although useful for genetical studies, should be used with caution specially when basing homology on cross-hybridization and fragment sizes.