ELISA: not a name, but a test.

Introduction: In our classroom we will simulate the spread of a "disease" by transfering "bodily fluids". After we have spread the "disease" we will then test to see who has it using enzyme-linked immunosorbent assay or ELISA. ELISA is used to test people for diseases such as HIV and to determine whether or not someone is pregnant. When someone has a disease their body has an immune response sending millions of antibodies. These antibodies locate and attach to their target antigens. Once the antibodies are attached other cells know to kill the harmful antigens. Antibodies are very specific and each one only recognizes one antigen. In the ELISA test secondary antibodies are linked to enzymes. The secondary antibodies attach to the primary antibodies that have already located the harmful antigens. In our kit the secondary antibodies are linked to horseradish peroxidase or HRP. When HRP comes in contact with hydrogen peroxide (the chemical added to the sample), HRP catalyzes the oxidation of TMB. The oxidation creates a blue product that turns the sample blue. If our sample turns blue then it means the "disease" is present. If it's clear then the "disease" is not present. After we find who is "diseased" we will attempt to trace it back to the original culprits.

Under the Sea

Introduction/Procedure: Before biotech and genomics, scientists studied animal behavior, habitat, and diet to distinguish the relationships between species. These conclusions were often wrong because factors such as behavior do not show how genetically close two species are. Just like genomics is the study of genes, proteomics is the study of proteins. However, proteomics is much more complex than genomics. While genome is rather consistent, the proteome is differs from cell to cell. In addition, the proteome regularly changes through biochemical transactions. Organisms can have drastic differences in protein expression during different life cycles, different parts of the body, and different environmental conditions. Scientists belive that much of the noncoding DNA functions to highly regulate protein production. The main component separating us from other organisms is our complex proteomes, not our genes. In our lab (the sushi lab) we will be taking a sample of fish and use proteomics to find the relation between different species. The first step of our lab is to denature the proteins using heat and or chemicals. Next, we will run them on a protein gel. Once the gel is finished we will analyze the results. The more similar that the bands on the gel are, the more related the species are.

Mountain DNA

Introduction: Not only do humans have 46 chromosomes but also each mitochondrian contains DNA. This DNA (mt DNA) contains only 37 genes which are all involved in the creation of energy. If one of these genes is diseased it effects the persons ability to generate energy and therfore is often tired. It is believed that mitochondria (which is bacteria) was taken up by the primitive human. The mitochondria could then feed off the helathy environment and help humans survive. Since mitochondria is bacteria, it has a higher rate of mutation. MtDNA has helped scientists trace history of humans. They used mtDNA polymorphisms to create a "family tree" and discovered that all humans come from a common ancestor. It is estimated that humans came from one ancestor around 200,000 years ago. The common ancestor became known as the "mitochondrial Eve". It is a reference to the biblical story of Adam and Eve however, presents the idea that there was no male present. Scientists say there was no male present because mtDNA is it's passed on maternally, meaning the mother is the only contributor. MtDNA is also used in forensic biology especially when the DNA is extremely damaged. It has been used to identify unknown soldiers after the Vietnam war, identify the Romonov royal family, and determine the relationship of Neandertal reamins to modern humans. In our experiment we will be going through the same process as the last lab to discover whether or not we contain a certain gene in mtDNA.

My Biotech Expertise, Show I Carry the "Disease"

Introduction: DNA testing is a very helpful and useful procedure. As we did earlier in the year DNA testing can be used to find out whether or not a plant is genetically modified. Other uses include; identifying if someone has a certain disease, solving crimes with DNA at the scene, confirming relationships between family members and determining the paternity of a child. To test for the "disease" we must extract the DNA. The first step in DNA extraction is breaking open the membarne with a 95 degree water bath. Then to make sure our DNA is not killed, we use Instagene matrix beads to wipe out the DNAse.The next process, polymerase chain reaction, (PCR) replicates the DNA so we can identify whether or not the "disease" is present. PCR requires four ingredients, target DNA, nucleotides, the enzyme thermus aquaticus, and primers. The first step to PCR is applying heat so that the two DNA strands are seperated. Next, Taql and DNA polymerase are added along with two short lab made primers. The temperature is lowered and the primers bind by complementary base pairing. Taql DNA polymerase adds bases and creates a new strand. This process repeats and the DNA grows exponentially. The final process is gel electrophoresis. We must run a gel to analyze the results. There are three possibilities: 1. homozygous dominant-healthy 2. heterozygous-healthy 3. homozygous recessive-diseased!

Results/Observations: After running the gel we all hoped to not be the one who is found to have the "disease". After a day of waiting for our results the moment of truth rolled around and it was not pretty. I was found to be the only group member with the "disease" and with this realization came much emotional pain. I was homozygous recessive (2 diseased) making me "diseased". Of the remaining three members two were found to be heterozygous (1 diseased, 1 normal). Since the normal trait is dominant they turned out okay. The final member was homozygous dominant (2 normal) obviously making him not "diseased". Although I had to deal with the emotional pain I found the lab to be quite interesting and I learned a lot about the passing on of traits.

Green Beans With Banana Genes

Introduction: Genetically modified organisms are exactly what they sound like. They are organisms that have been tampered with to create similar organisms with a few changed characteristics. Although farmers and other groups have been tampering with the genes of crops for a long time, it is now possible to insert a specific gene of interest into another organism. GMO's are created by using T.I. plasmids that contain the gene of interest. The T.I. plasmids go through transformation and the agricultural bacteria replicates its DNA. This bacteria is put into a plant cell which will eventually be grown into a GM plant. To identify GMO's the cell wall must be mashed up to create a wounded target. Whatever is being tested is then put into a hot H2O bath that breaks open the cell membrane. Next, instagene matrix is used to kill DNAse. With DNAse out of the picture DNA is open game. The next process, polymerase chain reaction, (PCR) replicates the DNA so we can identify whether or not it is genetically modified. PCR requires four ingredients, target DNA, nucleotides, the enzyme thermus aquaticus, and primers. Once the DNA has been replicated plenty of times, it's time to find out the answer. Found in 85% of GMO's, T.I. plasmids are the target that will prove if the organism is genetically modified. A gel must be run and if the GM band shows up then the organism is genetically modified, and if the plant band appears it is not. As it should be, genetically modified food is a controversial topic because both sides have strong points. Those in favor of GMO's argue they improve nutritional value, can be mass produced, and selective breeding brings out the best in foods. Those who oppose  GMO's have found the negatives in this process. Amongst their concerns are moral values in regards to tinkering with evolution, allergic reactions, and the problems that accompany a monoculture of products. Do the positives outweigh the negatives? Since GMO's are fairly new technology it's almost impossible to be certain. Our lab will consist of bringing in a food product from the store and doing all of the steps above to discover whether or not that food is genetically modified. Since so much of today's food is genetically modified, I believe our food will also prove to be a GMO.

Results/Observations: Our final gel had a band in every single lane. Since the first lane, test food with plant primers appeared, we know that DNA extraction worked. The second lane was test food with GMO primers. The band appearing in this lane proved that our first test food was in fact GMO. The third and forth lanes had the same results but were just a different food. The fifth lane was a GMO positive control DNA with plant primers. The fact that a band appeared in this lane shows that PCR was successful. Lane six was a GMO positive control DNA with GMO primers which reinforced that PCR worked. Finally lane seven contained just a marker. The band in this lane proved that gel electrophoresis worked.

The Glowing Belly of a Sea Jelly

Introduction:
In this lab we will be completing a genetic transformation by taking the gene that codes for Green Fluorescent Protein (GFP) and inserting it into the DNA of bacteria. Green Fluorescent Protein is found in sea jellys and we will take it to create glowing bacteria. Bacteria contains plasmids which are circular pieces of DNA. These plasmids usually contain genes that code for traits in bacteria and have the ability to transfer back and forth between bacteria. We will be inserting pGLO, a plasmid that codes for the GFP gene into the E. coli cells. The new genetically modified E. coli cells will then glow. To get the pGLO plasmid through the cell membrane we will need to use a solution of CaCl2 and use heat shock. Then to get the newly transformed cells to grow we will provide nutrients and incubate them. This will eventually make the cells start to express their fluorescent gene.