The analysis of the IGS sequence indicated that the restriction enzyme BglII cleaved the PCR products into fragments that are useful tools for the detection of A.flavus and A. parasiticus [19 (link)].
Following amplification, the PCR products were digested with the restriction enzyme BglII (Roche, Germany). The reactions were performed in a total volume of 40 µL containing 15 units of enzyme, 4 µL of buffer, 15 µL of the PCR product, and ultrapure water up to 40 µL. The reaction mixture was incubated at 37 °C for 1 h. Then, digested fragments were separated by electrophoresis on a 2% w/v agarose gel at 80V for 2.5 h and visualized under UV transillumination. The sizes of the resulting DNA fragments were estimated in comparison with a commercial 100 bp DNA ladder.
Following amplification, the PCR products were digested with the restriction enzyme BglII (Roche, Germany). The reactions were performed in a total volume of 40 µL containing 15 units of enzyme, 4 µL of buffer, 15 µL of the PCR product, and ultrapure water up to 40 µL. The reaction mixture was incubated at 37 °C for 1 h. Then, digested fragments were separated by electrophoresis on a 2% w/v agarose gel at 80V for 2.5 h and visualized under UV transillumination. The sizes of the resulting DNA fragments were estimated in comparison with a commercial 100 bp DNA ladder.