GMOs
What does the acronym GMO stand for?
Genetically Modified Organism
What does the acronym PCR stand for?
Polymerase Chain Reaction
What is the first step of DNA extraction?
You use a mortar and pestel or other tools to crush your sample.
What tool do you use to load the gel?
A micropipette.
Can you use a micropipette without a tip?
NOOOOOOOOOOOO
List at least 2 other GMO screening markers.
Screening genetic elements like
CAMV35s, p35S, tNos, pat, or bar or event specific markers for the official GMOs like Mon810, Bt11, or GT7.
What purpose do you think the Primer Mix serves in PCR?
Primers are important because they will hybridize with the sampled DNA and define the region of the DNA that needs to be amplified
Where is DNA stored within your food product sample?
Within the nucleus of the cell, in this case the cells that make up the main food product of your sample.
What is the charge of DNA?
Negative
Where do you store PCR samples?
In the fridge/freezer.
List at least 2 other examples of the use of genetic technologies outside agriculture.
Personalized medicine, CRISPR, IVF, Crime Scene Investigations, Species identification, Medicine & Research (knockout animal models, viruses, etc.)
What kind of promoter sequence do you anticipate for your positive control to express during PCR?
CAMV35s
What is the purpose of the DNA-EZ™ Lysis Solution?
Lysis buffer will break open the cells to expose organelles that contain DNA.
What is the purpose of the DNA ladder?
Used as a "ruler" to measure the distance between bp and draw conclusions from the samples, using established bp lengths as a guide.
Why is it necessary to balance your centrifuge before starting your sample? How do you balance it?
To avoid damage to samples (they will fly off and your sample will contaminate the entire centrifuge) and the device.
Placing a PCR tube on opposite ends of your sample(s).
If you were to test the presence of GMOs in a vegetable oil made with GMOs, what do you think the results would be? Why?
The refining process removes nearly all non-
triglyceride ingredients, therefore it is reported that there is no or a significantly small amount of protein or DNA remaining in vegetable oil.
At what temperature do you denature DNA?
~94°C
How long do you incubate your food mix in the DNA-EZ™ Lysis Solution and at what temperature?
Incubation Time: 5 mins
Temperature: 95 C
Why are controls necessary when visualizing data using gel electrophoresis?
Controls are needed for the validity of data.
Given a 2-20uL pipette, how would you dispense 75.5 uL of a solution? Provide a brief explanation & volume readouts for your measurements.
TA discretion.
You are walking across the streets of New York when you find a small piece of turtle shell near the sewer. You decide to bring this sample back to El Paso to a lab where your team is trying to decipher if this shell belonged to one of the teenage mutant ninja turtles, what are some of the steps your team would need to take to perform an experiment to verify that this turtle is a GMO?
You would do some preliminary research to find existing GMO models of turtles, this would help identify potential genetic markers. Furthermore, access to turtle genomes can also provide some references for novel sequences, which in turn, could be new GMO markers.
What would happen if the PCR step in your MiniOne thermocycler isn’t set to the right parameters? How would this affect your DNA copies?
PCR cycles have been optimized to ensure that efficient amplification takes place.
However, deviations to these recommendations can result in non-specific bands, no band (faint band), or smeared bands.
Do you think that processed foods vs. fresh produce are more suitbale for extraction? Which one do you think is best?
Processed foods are easier and more suitable for extraction, these products are preprocessed and homogenized making extraction of the right product successful.
Fresh produce might be difficult, one must strategize what portion of the food to use for extraction. For example, in strawberries the leafy part is used for an okay extraction.
What principles does gel electrophoresis rely upon to permit separation of DNA fragments into bands?
What would happen if the PCR step in your MiniOne thermocycler isn’t set to the right parameters? How do you think this would affect your gel electrophoresis results?
Some possible outcomes include . . .
* Fuzzy bands
* Primer bands below ladder markers
* No bands (failed PCR)