GMO Questions Week 1 (1)

Name Andrea Franco Group # 2 Date 11/11/2018 LABORATORY REPORT FORM Week One PCR AND GEL ELECTROPHORESIS LAB All answers to the following questions are to be typed in the spaces below, printed, and turned in at the beginning of the next lab meeting. Students should work individually to answer the following questions and in creating the figures. How can you test a food to find out if it contains material derived from a genetically modified organism (GMO)? The ELISA test can be used to detect the proteins in the GMO sample you’re testing. The PCR test can be used to expand GMO DNA sequence. The usage of these tests depend on whether the food sample is highly processed or not. In what organelles is plant DNA located? Plant DNA can be found in the three places. These include the mitochondria (powerhouse of cell), chloroplasts (organelles that work with photosynthesis), and nucleus (location of cellular information). 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.? If the temperature is too high or the DNA is tampered with, the plant DNA will be destroyed. What molecules are present in the cell that might interfere with DNA extraction? The DNase enzyme can degenerate DNA. Metallic ions are coenzymes that help the degradation of DNA. In plant cells, the cellulose wall can block the extraction of DNA. Why do you also perform analysis on food that is known to be a non-GMO food control? The analysis is performed to verify the non-GMO food sample has no unwanted particles on it. The non-GMO sample’s pattern from electrophoresis is used as a reference for foods that are genetically modified. Why are you performing two PCR reactions on each DNA sample? The first reaction was a control test to show that the extracted plant DNA was done using primers to a universal plant DNA sequence. The second reaction helped determine GMO sequence. What is the purpose of the GMO-positive control DNA? The purpose of the GMO-positive control is to see if the PCR successfully worked. A non-GMO result could be determined by a band in the showing up in the positive result sample. If 200 base pair band didn’t appear, it would mean the PCR didn’t occur. What does PCR allow you to do with DNA? PCR allows you to make multiple copies of a single DNA fragment. What components do you need to perform PCR? The components you need are a fragment of DNA, polymerase (enzymes) such as Taq polymerase, nucleotides such as dNTP, primers, and a cofactor which is a metallic ion. What is in the master mix and why do you need each component? The master mix includes the nucleotides, enzyme, and buffer needed for the PCR reaction. You need the cofactor to stabilize the polymerase (enzyme) reaction. This makes sure the enzyme doesn’t denature. The dNTP acts as an extension for strands of DNA. What steps make up a PCR cycle, and what happens at each step? The first step is denaturing with heat which splits the double-stranded DNA making a single strand. Next, annealing is done. Annealing decreases the temperature in the PCR reaction to combine the primer and single DNA strand. Afterwards, elongation raises the temperature to begin copying the DNA. 12