Multiplex PCR

The multiplex PCR assay was used to detect the genes for enzymes involved in flavor-forming pathways. The primers were designed manually by using the Primer-BLAST tool of National Center for Biotechnology Information (NCBI, USA) and were commercially obtained (Bioneer, Korea). Amplification was performed on a PCR system in a 20 μl reaction mixture consisting of i-Taq 2X PCR Master Mix Solution (iNtRON, Korea) with one colony of each streaking plate. The following PCR program was used for 30 cycles: 30 sec at 95°C, 20 sec at 41- 43°C, and 2.2 min at 72°C. The amplified products of each group were run on 1.5% agarose in 1× TAE buffer at 70 v for 40 min.