This study constructed a wheat-specific primer and a probe using the internal transcribed spacer (ITS). 2 regions of wheat (Triticum aestivum) and real-time PCR conditions were established. The calibration curve showed a slope of -3.356, a correlation coefficient of 0.998, and an amplification efficiency of 98.589%. Experiments were carried out on the rice flour mixed with 50%, 10%, 1%, 0.1%, 0.01%, and 0.001% of wheat. The result showed that it was possible to detect samples mixed with up to 0.01% of wheat. As a result of checking, the wheat detection potential of rice, 34 processed foods, and seven processed foods was ascertained. The real-time PCR method using the wheatspecific primer and probe developed in this study can be used to identify the authenticity of the raw materials, such as the incorrect indication of the raw materials utilized and the unintended mixing of wheat during the manufacturing process.

In this study, seven oligonucleotides primers were shown to generate the shared loci, specific loci, unique shared loci to each species and shared loci by the three species which could be obviously calculated. Euclidean genetic distances within- and between-species were also calculated by complete linkage method with the sustenance of the hierarchical dendrogram program Systat version 13. The genomic DNA isolated from herring (Clupea pallasii), Korean anchovy (Coilia nasus) and large-eyed herring (Harengula zunashi), respectively, in the Yellow Sea, were amplified several times by PCR reaction. The hierarchical dendrogram shows three chief branches: cluster 1 (PALLASII 01, 02, 03, 04, 06 and 07), cluster 2 (NASUS 08, 09, 10, 11, 12, 13 and 14), and cluster 3 (ZUNASHI 15, 16, 17, 18, 19, 20, 21 and PALLASII 05). In three clupeid species, the shortest genetic distance displaying significant molecular difference was between individual PALLASII no. 03 and PALLASII no. 02 (0.018). Individual no. 06 of PALLASII was most distantly related to NASUS no. 11 (genetic distance = 0.318). Individuals from herring (C. pallasii) species (0.920) exhibited higher bandsharing values than did individuals from Korean anchovy (C. nasus) species (0.872) (P<0.05). As a result, this PCR analysis generated on the genetic data displayed that the herring (C. pallasii) species was widely separated from Korean anchovy (C. nasus) species. Reversely, individuals of Korean anchovy (C. nasus) species were a little closely related to those of large-eyed herring (H. zunashi) species.