Add 50uL of Test Sample and NGMO sample to its designated tube that contains 500uL of Instamix. Both tubes are placed in 95°C water bath for 5 minutes and are then spun in the micro-centrifuge for 10 minutes. To set up for the PCR reactions, a Master Mix (MM), which contains DNA Primers, DNA Polymerase, Complementary base pairs, and a buffer, is required. For the first reaction, mix 20uL of NGMO food with 20uL of Plant MM (PMM). For the second reaction, mix 20uL of NGMO food with Genetically Modified MM (GMM). Repeat these step for test food sample and GMO sample with the two Master Mix's. There are 6 reactions in total. The 2 NGMO food sample will serve as a negative control to monitor any contamination in the sample. The 2 GMO food sample will serve as a positive control to ensure the Master Mix is functioning properly. The 6 solutions are placed in a freezer to allow the reaction to go through. To set up for electrophoresis, the gel is placed in the chamber and filled with buffer. Add 10uL of Orange loading dye into each sample. Load 15uL of solution from each of the 6 amplified DNA samples into the holes of the gel alongside with 15uL of molecular weight ruler on the 7th hole. Run the gel at 100V for 45 minutes. Using Gel Doc Imager, the photo of the gel get taken. Using the migration distance of the molecular ruler, an equation can be obtained through Microsoft Excel by graphing the standard curve. By measuring the migration distance of each band and utilizing the equation obtained, the length of base pair can be calculated(1).
Results.
This experiment was conducted with five different samples of test food, including soya flour, wheat, corn, rice, and corn meal. Using Gel Electrophoresis, high-quality DNA migrated from its original position due to the electrical current. The photos of the gel obtained by Gel Doc Imager provided a clear image of which band is present in every lane. For all 5 test food samples, no bands are present in lane 2.