BIOL 1500- D05

Week 1: GMO Lab Assignment

1. How can you test a food to find out if it contains material derived from genetically

modified organism (GMO)?

There are two main methods. The first being ELISA (enzyme-linked immunosorbent assay),

which detects proteins produced specifically from GM crops. It is not, however, useful in testing

highly processed foods as the proteins may have been destroyed. The second method is PCR

(polymerase chain reaction). This amplifies the genetically modified DNA and analyzes it with

gel electrophoresis. Since DNA is more resistant than proteins, it can be extracted from highly

processed foods.

2. In what organelles is plant DNA located?

Plant DNA is found in the nucleus, mitochondria, and chloroplast.

3. Many foods containing GM crops are highly processed. Can you suggest how DNA

from whole plants may differ from that extracted from processed foods, e.g., corn

chips, cornmeal, etc.?

Highly processed food may denote heating or high temperatures, which could allow for the

denaturing of DNA, thus leading to fragmentation. The genetic makeup is different from less

processed foods.
 4. What molecules are present in the cell that might interfere with DNA extraction?

Enzymes (DNase) can degrade DNA. Cofactors (metal ions) and coenzymes may degrade DNA.

Though salt and magnesium ions are needed for Taq DNA polymerase to perform optimally, too

much or too little may have an effect.

5. Why do you also perform analysis on food that is known to be a non-GMO food

control?

Some samples may have been contaminated, and it would check for that. It would also be

performed as a comparison; it can show what a sample looks like in the absence of GMO’s.

6. Why are you performing two PCR reactions on each DNA sample?

The first reaction shows the extraction of plant DNA through primers to a universal plant DNA

sequence. The second reaction identifies the GMO target sequence.

7. What is the purpose of the GMO-positive control DNA?

It is a confirmation that the PCR reaction worked. Positive controls produce a positive result. If

there is no band in the test sample, the test is most likely non-GMO. If there is no 200 base pair

band in the positive control, it is assumed the PCR reaction did not work.

8. What does PCR allow you to do with DNA?

PCR amplifies a segment of DNA that can be a few hundred base pairs in length, and copies it.

This allows for many copies and more opportunities to perform analysis.
 9. What components to do you need to perform PCR?

The DNA template, DNA Polymerase, oligonucleotide primer, nucleotides and master mix

(which contains buffer).

10. What is in the master mix and why do you need each component?

A polymerase buffer (which has the ions essential for catalysis), dNTP (contains adenine,

thymine, guanine, cytosine), Primers (to serve as a start/stop point in amplifying the target region

of DNA to be copied), and DNA Polymerase (initiates the addition of nucleotides).

11. What steps make up a PCR cycle, and what happens at each step?

There are three main steps. In Denaturation, the double stranded DNA is heated and melted, and

the single strands can be replicated. A primer is necessary in the next part as it starts and stops

points for amplifying the target regions of the DNA that needs to be copied. In Annealing, the

primer sticks to the single strands of the DNA and start the signal for replication. Hybridization

is the binding of the oligonucleotide primer to the template DNA. In Extension, the DNA

polymerase allows for the starting of DNA replication.