Identification of Genetically Modified Food (GMO) by DNA, PCR and Gel Electrophoresis (2685)

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QuestionQuestion

Write a full lab protocol report on DNA, PCR and Gel Electrophoresis

Abstract:
-Concise summary of the entire experiment, must have at least one sentence for each part of the manuscript: Introduction, Methods, Results and Discussion.
-Must have a clear hypothesis and conclusion
-150-200 words
- 3 or 4 keywords

Introduction: Need to include at least 2 primary sources (=journal articles). A great paper will have a lot more than 2 primary sources.
-Background on GMO? Some of the following issues may be addressed:

• How are plants genetically modified?
• Why are plants modified?
• What modifications exist?
• How are the modification detected?
• Where are they found, consumed or how are they labeled?
• Pros and cons of GMOs

-Brief description of the experimental methodology.
-What are target genes for the plant and GMO PCRs?
-Objective and Hypothesis: Do grocery products contain genetically modified organisms? Which unknown sample was a potential GMO? What is your hypothesis?
*Examples from the literature must be given- don’t just list details off the Power Points, or other materials provided in blackboard.

Methods:
-Summary of the methods that were used: DNA extraction and PCR, and Gel electrophoresis.
*Protocol specific details, such as PCR cycle times, gel voltages, centrifuge speeds, temperatures, incubation times , etc. should be included. These are all things that are absolutely critical for a scientist to know so that they can repeat your experiment. If they change these things, the experiment will come out differently!
*Details such as volumes, tubes, vortexing, etc. SHOULD NOT be included! These are things an experienced scientist can work out for themselves. If you are confused about what should and should not be, look at examples from the primary literature!

Results:
-Description of the PCR results (plant and GMO) for all your samples. You must electronically label ALL bands of the MW ladder, and state all bands for your group (presence or absence) and size in the text.
*No interpretation of the results- just state findings

Discussion:
What do the results mean? Explain experimental errors and potential sources of negative results.

-Include the roles of the positive and negative controls, and the relevance of plant and GMO PCR’s in determining the GMO status of your test food.
- What can you conclude about the test samples? What else could be done to complement your findings?
-Do these results support the results of other scientists (i.e. do your results support or disagree with the literature)
-What are some reasonable future experiments- how can you take this specific set of experiments a step further (hint: look to the literature to see what others have done)
*Need at least 2 primary sources in your discussion

Appendix
- Figures! You need to have the gel, but feel free to include other fun figures, such as pictures of your test samples, etc.

References:
-At least four sources from the primary scientific literature.
- Also cite lecture material when appropriate.
-Follow correct citation format!

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Identification of Genetically Modified Food (GMO) by isolation of genomic DNA, PCR amplification and gel electrophoresis.

Abstract
Genetically Modified Organisms (GMO) are organisms whose genomic DNA sequences have been engineered with an introduction of foreign DNA. GMO plants are made to introduce a specific phenotype of superior trait such as high yield, tolerance and resistance to pests. But they have not been widely approved for source of food because of the apprehensions of the obvious idiosyncratic side effects that are unforeseen and unpredictable. It is important to identify a given food material has a GMO source. This can be done using PCR, with primers designed to amplify the foreign sequences commonly used in GMO. In the current experiment PCR screening for GMO was performed from banana. DNA was isolated from banana using InstaGene matrix. This DNA was used in PCR using GMO specific and plant specific master mixes and after PCR, the products were separated on 3% agarose gels, in TAE. Positive control specific to GMO and negative controls were also employed in PCR. We achieved partial success with the experimental setup. Bands were observed in positive controls indicating that the PCR was performed well. Absence of bands in negative control indicated that the protocol was contamination free. However, no bands were observed in the DNA from the test sample, possibly due to low yield in isolation or due to inhibitors carrying over during the DNA isolation which could inhibit PCR. Further reactions are necessary to confirm the success of the protocol as well as to determine whether the test plant is GMO or not.

Key words: GMO, Genetically modified organisms, PCR, DNA extraction, Agarose gel electrophoresis

Introduction
It all started with the science of genetics, where traditional plant breeders crossed superior genotypes. This was done to create genotype combinations with desirable traits in plants (University of Nebraska, 2001). Discovered in 1983, a new type of modification, not limited to the results of crossing is genetic engineering (Davison & Bertheau, 2007). Unlike traditional breeding, this type of engineering allows adding a foreign gene to the genome of an organism and does not transfer undesirable traits. In addition, traits from any living organism can be transferred into a plant, since there is no sexual barrier. The process: includes extracting the DNA, followed by gene cloning which separates the gene of interest. The gene that was cloned must be then designed to work if...
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