Improvement in group identification of dojo loach, Misgurnus anguillicaudatus, using PCR-restriction fragment length polymorphism

In most Japanese populations of dojo loach (Misgurnus anguillicaudatus), gonochoristic diploids genetically diversified groups (A and B, further subdivided into B1 B2) are present, whereas unisexual clonal lineages inhabit certain localities in the Hokkaido Ishikawa Prefectures Japan. Through a series genetic studies including DNA markers, loaches were deemed to originate from hybridization event(s) between A groups. However, combined analyses with other markers needed identify each group. this study, we improved PCR-restriction fragment length polymorphism (RFLP) analysis recombination activating gene 1 (RAG1) using digestion two restriction enzymes, PvuII StuI. The RAG1-RFLP showed different patterns for group: fragments (245 198 bp) group A, three (198, 147, 98 B1, single (443 B2. exhibited four (245, 198, derived both B1. Moreover, able detect hybrid genotypes × B2 B2). Thus, allowed quick accurate identification loaches.