Charomid cloning vectors meet the pedipalpal chelae: single-locus minisatellite DNA probes for paternity assignment in the harlequin beetle-riding pseudoscorpion

We describe the first application of the charomid-cloning method for developing single-locus minisatellite DNA probes in a terrestrial arthropod. From a genomic library of the neotropical pseudoscorpion, Cordylochernes scorpioides , we have isolated two probes with heterozygosities exceeding 95%. These probes yielded single-locus patterns after only low stringency washing and in the absence of genomic competitor DNA. Analysis of three pedigrees indicated germline stability and showed no evidence of linkage between the loci. Patterns of allelic transmission generally conformed closely to Mendelian expectations but large offspring numbers did enable detection of one example of significant bias in allele inheritance. Two test cases are presented to illustrate the clarity and power with which these probes can establish paternity: (i) a female mated to three unrelated males, and (ii) a female mated to two of her brothers. In both cases, a single probe could be used to assign the paternity of all offspring.