*2.8. PCR Assay for A. flavus Detection*

′ ′ ′ ′ ′ ′ ′ ′ For specific detection of *A. flavus*, 64 PCR reaction contained 2 µL of the extracted DNA and 23 µm PCR mix containing 11.5 µL Taq DNA polymerase (Jena, Germany), 5 mM of two specific primers for *A. flavus aflP* (F-5′ -CATGCTCCATCATGGTGACT-3′ ), (R-5′ CCGCCGCTTTGATCTAGG-3′ ) [30], *aflA* (F-5′ -GGTGGT GAAGAAGTCTATCTAAGG-3′ ), and (R-5′AAGGCATAAAGGGTGTGGAG-3′ ) [31]. PCR thermal cycler program was adjusted as follows: 7 min at 94 ◦C tracked by 40 amplification cycles at 94 ◦C for 30 s, annealing temperature 62 for 30 s, 72 ◦C for 30 s, and then 72 ◦C for 3 min for the final extension. The amplified DNA was separated via 2% agarose gel electrophoresis containing ethidium

bromide at 90 V for 30 min. Agarose gel was detected in UV transilluminator light via Gel Documentation System (Uvitec, Cambridge, UK).
