Presentation gmo (2)


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  • 35S promoter of the cauliflower mosaic virus (CaMV 35S)NopalineSynthase(NOS) gene of Agrobacteriumtumefaciens.Duplex vs. SimplexThere was two different types of procedures in order to identify if a fruit is genetically modified- PCR that is for gene or ELISA- that is for proteins. Actually, a promoter is involved in RNA transcription from a DNA sequence. It regulates the binding of RNA polymerase and the rate at which RNA is transcribed.Promoters are specific sequences of DNA where polymerase initiates transcription. The terminator is a sequence that signals the end of transcription.For translation a AUG is the start codon, and translation stops at the STOP codon.A transgene is any foreign DNA that has been inserted into the genome of an organism, typically using transposable elements.
  • Explain 2 grams of each sample.
  • Red- primersforterminator(NOS) NopalineSynthaseofAgrobacteriumtumefasiensandpromoter (CamV35-Cauliflowermosaic virus)Greenisforthephotsystem gene identification-Indentifyifthesampoleis a productof a plant.
  • Red iswho determineiftheproductis a GMO.
  • Presentation gmo (2)

    1. 1. Presence of CaMV35S and NOS in Carica Papaya, Glycine Max, Triticum spp. and Zeamays<br />Valeria Rivera-Torres<br />Carlos Santos-Perez<br />Juan Feliciano-Figueroa<br />RISE Students<br />Mentor: Dr. Belinda Roman-Aviles <br />
    2. 2. Introduction<br />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. <br />A GM plant contains a promoter and a terminator in order to create the modification in the DNA sequence of the organism. <br />Promoter-35S <br />Terminator- NOS <br />PCR vs. ELISA <br />
    3. 3. Objectives<br />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.<br />Carica papaya (Papaya)<br />Glycine max (Soy milk)<br />Triticum spp. (Wheat)<br />Zeamays (Whole Corn)<br />
    4. 4. Hypothesis<br />It is expected that Glycine max and Zeamayswill be Genetically modified whether Carica papaya and Triticum spp. will not be. <br />
    5. 5. Methodology<br />
    6. 6. Extracting DNA<br />Mortar and Pestle Technique<br />Grind with pestle for at least 2 min to form a slurry.<br />Instagene® (BioRad®)<br />Lysis the cells from this sample<br />Precipitate the DNA<br />Place the samples on the hot plate for 5 minutes at 37°C.<br />Denature Proteins<br />
    7. 7. Polymerase Chain Reaction (PCR)<br />Each tube contain the respective master mix<br />Mastermix- green<br />Mastermix-red <br />Each eppendorff tube contains: <br />NGMO PMM (green)<br />NGMO GMM (red)<br />GMO + PMMM (green)<br />GMO + GMM (red)<br />Carica Papaya PMM (green)<br />CaricaPapay GMM (red)<br />Glycine max PMM (green)<br />Glycine max GMM (red)<br />Each tube contain the respective master mix<br />Mastermix- green<br />Mastermix-red <br />Each eppendorff tube contains: <br />NGMO PMM (green)<br />NGMO GMM (red)<br />GMO + PMMM (green)<br />GMO + GMM (red)<br />Whole corn PMM (green)<br />Whole corn GMM (red)<br />Wheat PMM (green)<br />Wheat GMM (red)<br />
    8. 8. Electrophoresis <br />Make the agarose gel<br />3% of Agaraose<br />Add 10 microliters of Orange G loading dye (LD) to each sample and mix well.<br />Load 20 microliters of the molecular weight ruler and 20 micro liters of each sample into your gel. <br />Ethidium Bromide <br />Reasons for the addition:<br />Orange G Loading Dye<br />Molecular Weight Ruler<br />
    9. 9. Data- Gel Electrophoresis<br />
    10. 10. Carlos<br />
    11. 11. Valeria<br />
    12. 12. Juan<br />
    13. 13. Conclusion <br />The hypothesis can not be accepted or rejected. <br />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. <br />
    14. 14. Future Plans <br />The researchers will make a repetition of the same process in order to improve the results.<br />This research can be apply to the agriculture in Puerto Rico in order to identify which of the Puerto Rican products are GMOs. <br />Interdisciplinary contribution to the Puerto Rican community. <br />The effects of GMOs present in the Puerto Rican diet. <br />