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Frank Soto
5/6/2011
 Background
 Genes to identify: 35S promoter, NOS terminator, and
PSII
 Methodology
a) DNA Extraction
b) DNA Amplificat...
 GMO’s are organisms that have been modified through
genetic engineering.
 In the United States, under guidelines issued...
Genes
Structure of a gene
 Steps for genetic
modification of a
crop:
a) First, find the
ideal protein to
express.
b) Isolation of the
gene of inter...
 35S promoter from the cauliflower mosaic
virus (CaMV 35S)
 Nopaline synthase (NOS) terminator from
Agrobacterium Tumefa...
 Our hypothesis is that the genes 35S and NOS
will appear in the gel. The 35S gene will be
found at 203bp and NOS at 225b...
 DNA Extraction
a) Weighing and
Grinding of samples
b) Pipette 50µL of Slurry
into tube containing
Instagene matrix.
c) F...
 PCR
a) 20µL of indicated
master mix is
dispensed to each
of the samples.
b) 20µL of DNA
samples are
dispensed to their
c...
 Electrophoresis
a) The gel used was a 3% Agarose
gel.
b) To prepare for electrophoresis,
10µL of Orange Loading dye was
...
Frank’s Gel
Lester’s Gel
 One can conclude that the maize was
genetically modified, while the soymilk,
papaya, and corn chips are inconclusive.
 ...
Investigation of genetic modification in maize and soymilk
Investigation of genetic modification in maize and soymilk
Investigation of genetic modification in maize and soymilk
Investigation of genetic modification in maize and soymilk
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Investigation of genetic modification in maize and soymilk

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Investigation of genetic modification in maize and soymilk

  1. 1. Frank Soto 5/6/2011
  2. 2.  Background  Genes to identify: 35S promoter, NOS terminator, and PSII  Methodology a) DNA Extraction b) DNA Amplification c) Electrophoresis of DNA  Analysis of Results  Conclusion
  3. 3.  GMO’s are organisms that have been modified through genetic engineering.  In the United States, under guidelines issued by USDA's 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.
  4. 4. Genes Structure of a gene
  5. 5.  Steps for genetic modification of a crop: a) First, find the ideal protein to express. b) Isolation of the gene of interest. c) Insertion of the ideal promoter and terminator d) Back cross to highest-yielding plants
  6. 6.  35S promoter from the cauliflower mosaic virus (CaMV 35S)  Nopaline synthase (NOS) terminator from Agrobacterium Tumefaciens  Photosystem II (PSII)
  7. 7.  Our hypothesis is that 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.
  8. 8.  DNA Extraction a) Weighing and Grinding of samples b) Pipette 50µL of Slurry into tube containing Instagene matrix. c) Flick tube and place in a 95 degrees celsius water bath for 5 min. d) Place tubes in microcentrifuge for 5 min. Non-GMO Test food Test food 50µL of sample slurry 500µL of Instagene matrix 50µL of sample slurry 500µL of Instagene matrix 50µL of sample slurry 500µL of Instagene matrix
  9. 9.  PCR a) 20µL of indicated master mix is dispensed to each of the samples. b) 20µL of DNA samples are dispensed to their corresponding tubers. c) Once the samples have been dispensed, the tubes are put in the thermal cycler. Non-GMO PMM Non-GMO GMM GMO (+) PMM GMO (+) GMM Test PMM Test GMM Test PMM Test GMM PMM= Plant molecular marker GMM= Genetic molecular marker.
  10. 10.  Electrophoresis a) The gel used was a 3% Agarose gel. b) To prepare for electrophoresis, 10µL of Orange Loading dye was deposited into each of the samples. c) 20µL of Molecular weight ruler(mwr) was deposited to wells to wells 1 and 10. d) 20µL of samples were dispensed to wells 2-9. e) Electrophoresis was run for 30min. at 100V. f) After electrophoresis, the gel was stained with ethidium bromide pads for 20min. 1 Molecular weight Ruler 2 Non-GMO PMM 3 Non-GMO GMM 4 GMO (+) PMM 5 GMO (+) GMM 6 Test PMM 7 Test GMM 8 Test PMM 9 Test GMM 10 Molecular Weight Ruler
  11. 11. Frank’s Gel
  12. 12. Lester’s Gel
  13. 13.  One can conclude that the maize was genetically modified, while the soymilk, papaya, and corn chips are inconclusive.  The experiment must be repeated because the results were affected by human errors, such as bad pipetting technique.

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