1. Presence of CaMV35S and NOS in Carica Papaya, Glycine Max, Triticum spp. and Zeamays Valeria Rivera-Torres Carlos Santos-Perez Juan Feliciano-Figueroa RISE Students Mentor: Dr. Belinda Roman-Aviles

2. Introduction Genetically Modified Organism (GMO) refers to animals, plants and microorganisms that are inserted with a foreign gene or genes, that partially or completely change their metabolic pathway. A GM plant contains a promoter and a terminator in order to create the modification in the DNA sequence of the organism. Promoter-35S Terminator- NOS PCR vs. ELISA

3. Objectives This experiment focuses on detecting the presence of genetically modified organisms using food items to extract DNA, amplify using PCR and run a gel electrophoresis. Carica papaya (Papaya) Glycine max (Soy milk) Triticum spp. (Wheat) Zeamays (Whole Corn)

4. Hypothesis It is expected that Glycine max and Zeamayswill be Genetically modified whether Carica papaya and Triticum spp. will not be.

5. Methodology

6. Extracting DNA Mortar and Pestle Technique Grind with pestle for at least 2 min to form a slurry. Instagene® (BioRad®) Lysis the cells from this sample Precipitate the DNA Place the samples on the hot plate for 5 minutes at 37°C. Denature Proteins

7. Polymerase Chain Reaction (PCR) Each tube contain the respective master mix Mastermix- green Mastermix-red Each eppendorff tube contains: NGMO PMM (green) NGMO GMM (red) GMO + PMMM (green) GMO + GMM (red) Carica Papaya PMM (green) CaricaPapay GMM (red) Glycine max PMM (green) Glycine max GMM (red) Each tube contain the respective master mix Mastermix- green Mastermix-red Each eppendorff tube contains: NGMO PMM (green) NGMO GMM (red) GMO + PMMM (green) GMO + GMM (red) Whole corn PMM (green) Whole corn GMM (red) Wheat PMM (green) Wheat GMM (red)

8. Electrophoresis Make the agarose gel 3% of Agaraose Add 10 microliters of Orange G loading dye (LD) to each sample and mix well. Load 20 microliters of the molecular weight ruler and 20 micro liters of each sample into your gel. Ethidium Bromide Reasons for the addition: Orange G Loading Dye Molecular Weight Ruler

9. Data- Gel Electrophoresis

10. Carlos

11. Valeria

12. Juan

13. Conclusion The hypothesis can not be accepted or rejected. Human errors are involved and affected the results of the investigation. Pipetting, concentrated DNA, lots of primer, evaporation, and contamination are among the possible errors that occurred during this experiment.

14. Future Plans The researchers will make a repetition of the same process in order to improve the results. This research can be apply to the agriculture in Puerto Rico in order to identify which of the Puerto Rican products are GMOs. Interdisciplinary contribution to the Puerto Rican community. The effects of GMOs present in the Puerto Rican diet.

15. References Cantamutto, M., & Poverene, M. (2007). Genetically modified sunflower release: Opportunities and risks. Fields Crops Research, 101, 133-134. Jiao, Z., Deng, J., Li, G., & Cai, Z. (2010). Study on the compositional differences between transgenic and non-transgenic papaya(Carica papaya L.). Journal of Food Composition and Analysis, 23, 640-647. Scipioni, A., Saccarola, G., Arena, F., & Alberto, S. (2008). Strategies to assure the abscence of GMO in food products application process in a confectionery firm. Food Control, 16, 569-578. Terzi, V., Malnati, M., Barbanera, M., Stanca, A., & Faccioli, P. (2003). Development of analytical systems based on real-time PCR for Trititcum species- specific detection and quatitation of bread wheat contamination in semolina and pasta. Journal of Cereal Science, 38, 87-94. BioRad® Manual of: Biotechnology Explorer™ GMO Investigator™ Kit