Is My Strawberry a Frankenfood?: GMO Foods Lab

Introduction
a) Background- GMO's are genetically modified organisms, and can be used in various scientific settings, often in agriculture, such as in the development of making genetically modified foods, that could be resistant to disease or frost or having a higher amount of vitamins in it. GMO's are made by taking a plasmid with the gene of interest (a tumor inducing plasmid) and putting it into agro bacterium. Then we will place the agro bacterium in a plant cell. The plasmic will take the DNA and will transcribe it, creating a genetically modified plant. GMO's are identified throught two techniques, ELISA and PCR. We are using PCR in  this lab, where we identify sequences of DNA that have been inserted into the GM plant. DNA fragments can be isolated from highly processed foods. GMO's are controversial because some people identify genetically modified organisms as "unnatural" and have the potential to create organisms that will be resistant to normal toxins. Many also worry about potential allergic reactions to GMO's and think that organic foods are better for the environment.
b) Purpose- The purpose of this lab is to take an organism and determine if it is genetically modified or not by comparing it to a known organic organism. The technique of PCR is used regularly in the scientific world today, for example in forensics and genetic studies.
c) Procedure- First we will mortar and pestal the cell of an organism to break down its cell wall. Then we will place the crushed cell in a 99 degree Celcius water bath to break down the nuclear and cell membranes. we will also have to add Instagene Matrix beads that kill DNAse enzymes, otherwise the DNAse will come into contact with the DNA because the cell wall is broken and will destroy it. Next, we will use the PCR process using two primers, one to target plant DNA (the control) and one to target GM DNA. Finally, we will use gel electrophoresis to visualize the gene of interest and determine the presence/absence of the desired PCR products.
d) Hypothesis- In this lab we are testing if the organism we bring in is genetically modified or not.
Procedure
On Day 1, we will label two tubes with Test Food 1 and Test Food 2. After we weighed out about 0.9 grams of non-GMO food (corn flour) and put it in the mortar, along with 10 milliliters of distilled water. We ground up this mixture for around two minutes and then followed by adding 5 milliliters of distilled water. Next we pipetted 50 microliters of the slurry into the tube containing InstaGene. After, we prepared our tube for our second test food, strawberries, using the same procedure. Then we flicked both tubes and placed the tubes in a 95 degree Celsius water bath for 5 minutes. We followed by placing the tubes in a centrifuge for 5 minutes and then we refrigerated the tubes over night. On Day 2, we labeled six PCR tubes. Tubes 1 and 2 contained 20 microliters of non-GMO food control DNA. Tubes 3 and 4 contained 20 microliters of test food DNA. Tubes 5 and 6 contained 20 microliters of GMO positive control DNA. After we labeled these tubes, we placed them in the foam float on ice. Then we added 20 microliters of plant master mix to tubes 1, 3, and 5 and 20 microliters of GMO master mix to tubes 2, 4, and 6. Finally we placed the PCR tubes in the thermal cycler.  On Day 3, we obtained our PCR tube from the thermal cycler and centrifuged it for three seconds. Next we pipetted 10 microliters of Orange G loading dye to test tubes 1-6. Then we loaded 20 microliters of each sample (in the tubes) into rows 1-6 of our gel. In row 7, Mr. Chugh loaded 20 microliters of PCR molecular weight ruler, to act as a control for our lab. Then we ran the gel for 3 minutes at 200 volts. The se stained in Fast Blast DNA stain. The next day, we observed our results.
Results
We concluded that our strawberries were genetically modified.

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Discussion
When observing our gel the next day, we found banding pattern in rows 3, 4, 5 and 6. Because there was a band in lane 4 of the gel, we can conclude that the strawberries we tested were genetically modified. One possible source of error for this lab was not mixing the loading dye well in each sample. Other sources of error could be that PCR or extraction could have been unsuccessful.