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We constructed a gramineous plant-specific primer set-matK 792F1GC and matK 979R1, targeting the maturase K (matK) gene, for detection of gramineous raw materials in processed food. For this, we used polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The primer set was evaluated using agarose gel electrophoresis and DGGE. Our results confirmed that the primer set was specific for gramineous plants. From our analysis, we confirmed that up to 6 types of gramineous plants can be simultaneously detected using PCR-DGGE. Results of a model test showed that by using this method, it was possible to detect wheat from a sample, if present more than 0.1 % (w/w). Thus, our method of PCR-DGGE with matK 792F1GC and matK 979R1 can be used to detect the presence of gramineous raw materials in processed food.