Fisheries Laboratories, Kinki University, 1Oshima Experiment Station, 2Oshima Fish Nursery Center, Kushimoto, Wakayama 649-3633, 3Shirahama Experiment Station, Shirahama, Wakayama 649-2211, 4Uragami Experiment Station, Nachikatsuura, Wakayama 649-5145, Japan
AFLP, amplified fragment length polymorphism screening, was conducted to identify sex characteristic DNA markers of cultured Pacific bluefin tuna (PBT), Thunnus orientalis. The AFLP screening of 4 females and 4 males with 64 primer combinations produced a total of 1038 scorable bands, of which 183 were polymorphic between individuals. Male characteristic fragments were confirmed with EcoRI+AGG and MseI+CAT primers by gel image analysis, which corresponded to 437 bp in 29 of 32 males and 3 of 32 females. Fragment detail was analyzed by electropherogram analysis and DNA quantification. The 437 bp fragment was detected in 30 individuals of average DNA concentration 0.231 ng/μL; while in females, 13 individuals were detected with average 0.080 ng/μL (P<0.01).