Genetic Modification in Maize, Papaya, Fritos® Corn Chips, and Natura® Soymilk
Frank SotoLester Rosario 5/6/2011
Overview Genes to identify: 35S promoter, NOS terminator, and PSII Methodologya) DNA Extractionb) DNA Amplificationc) Electrophoresis of DNA Analysis of Results Conclusion
GMO’s are organisms that have been modified through genetic engineering. In the United States, under guidelines issued by USDAs Animal and Plant Health Inspection Service, genetic engineering is defined as the genetic modification of organisms by recombinant DNA techniques (USDA, 2007) According to the USDA, crops have been genetically modified ever since 1996. These crops have all kind of modifications, such as: herbicide tolerance, pesticide tolerance, and bacterial resistance.
35S promoter from the cauliflower mosaic virus (CaMV 35S) Nopaline synthase (NOS) terminator from Agrobacterium tumefaciens Photosystem II (PSII)
The genes 35S and NOS will appear in the gel. The 35S gene will be found at 203bp and NOS at 225bp. We believe the corn will be identified as GMO while the soymilk won’t. In addition, both the corn chips and the papaya will be indentified as GMO.
DNA Extractiona) Weighing and Grinding of samples Non-GMO Maize Soymilkb) Pipette 50µL of Slurry into tube containing 50µL of 50µL of 50µL of Instagene matrix. sample sample sample slurry slurry slurryc) Flick tube and place 500µL of 500µL of 500µL of in a 95 degrees Instagene Instagene Instagene celsius water bath for matrix matrix matrix 5 min.d) Place tubes in microcentrifuge for 5 min.
PCR PMM= Plant molecular marker GMM= Genetic moleculara) 20µL of indicated master mix are marker. dispensed to each Non-GMO PMM of the samples. Non-GMO GMMb) 20µL of DNA samples are dispensed to their GMO (+) PMM corresponding tubes. GMO (+) GMMc) Once the samples Mai. Test PMM have been dispensed, the Mai. Test GMM tubes are put in the thermal cycler. Soy Test PMM Soy Test GMM
1 Molecular weight Ruler Electrophoresisa) The gel used was a 3% Agarose 2 Non-GMO PMM gel. 3 Non-GMO GMMb) To prepare for electrophoresis, 10µL of Orange Loading dye was deposited into each of the samples. 4 GMO (+) PMMc) 20µL of Molecular weight ruler(mwr) were deposited to wells 5 GMO (+) GMM to wells 1 and 10.d) 20µL of samples were dispensed 6 Mai. Test PMM to wells 2-9.e) Electrophoresis was run for 7 Mai. Test GMM 30min. at 100V.f) After electrophoresis, the gel was 8 Soy Test PMM stained with ethidium bromide pads for 20min. 9 Soy Test GMM 10 Molecular Weight Ruler