Antibody Testing Introduction

This experiment involves a mock outbreak of an infectious disease within our classroom. After the spreading is over we will use the process of ELISA to see who has the disease. Antibodies are Y shaped proteins that specifically bind to a certain foreign antigen. Millions of antibodies are made and destroyed every ten minutes. There are two types of antibodies primary and secondary. Primary antibodies bind specificially to the antigen, while secondary antibodies are usually made from a different species and bind to the primary antibodies. In the real world Enzyme linked immunosorbent assay is used for pregnancy tests, disease detection in animals and plants, as well as drug use etc.

Procedure:
Antigen is added to a microplate strip and incubates until bound/ washed with detergent.
Next primary antibody solution is added to the wells and allowed to bind/ than the unbound is washed away.
Next the secondary antibodiy is added and allowed to bind to the primary antibody, and than washed away.
Next a chromogenic enzyme is added and allows color to develop if there was binding earlier.

     

Proteanomics Conclusion

Procedure
I was actually absent for this lab but the procedure was pretty similar to most of our other labs. In this case samples of different fish, were cut off placed with a solution of buffer, incubated for a brief period and then heated up to denature the protein. Next gel electrophoresis is run on all of the proteins including, lanes set aside in order to prove or disprove if the first step worked. In our experiment the different lanes of the proteins all showed up except for that of the fly. Unfortunately enough flies have very little muscle tissue and in order to get enough tissue we needed about fifty flies rather than 3 or 4. From our results we were able to determine that the tuna and salmon were most closely related, followed by the shrimp and than the scallop, and than the fake crab, crab, and the fly didnt show up. Some sources of error could be not extracting enough protein, gaining some protein from particles floating in the air, mistaking certain bands because they were soo close together and etc.

Intro to Proteomics

Proteomics is the study of proteins mostly their structures and functions. The comparison of proteins from one species to another supports evolution and helps evolutionary scientists receive a better understanding of how closely a species is related. Proteonomics has risen mostly after the completion of the human genome project that took thirteen years, to be able to see the difference between genes and how they are expressed in certain situations. Recently the study of proteins has expanded to include the study of protein functions, interactions, locations, and modifications. It is hypothesized that the noncoding DNA regulates protein production and expression and because at 3 billion base pairs, we are much more complex than say a worm. The HUPO Human Proteome Organization was created specifically for cataloging all human proteins and function.
The term evolution states that organisms derived from simple single celled organisms and adapted individually to their environment. Mutations that were beneficial were passed on creating different species, that express different phenotypic traits. Muscle proteins consist mostly of myostotin and actin but individual species have different proteins for their specific types of movement.
This lab consists of three steps including 1) Denaturing the protein by heat shock as well as a detergent sodium dodecyl sulfate killing the protein tertiary and quaternary structures. The proteins then are put with a buffer and seperated using gel electrophoresis. Depending on the size of the different proteins from the aquatic animals it shall illustrate how closely our specimens are related. A blue tracking dye is added in order to see how far the proteins went.       

Mitochondrial DNA

In this lab we all had the same bands for this specific gene that we targeted, proving that PCR, and everthing in the lab went right. Possible sources of error could be, the cotton swab in the pipet not filtering enough giving DNA from the air, and some human sources of error could be going to low and taking some of the matrix beads therefore killing the DNA polymerase and stopping the DNA from being amplified.

Intro to Mitochondrial DNA

In Mendelian genetics the offspring of a mother and father should theoretically be 50% mom, 50% father, but in reality this is not true. The mother contributes more in her oocyte with additional organelles and extra DNA in the form of Mitochondria. Mitochondria contains an extra 37 genes and produces the energy the cell needs to function in the form of ATP.
Mitochondria is hypothesized to have originally come from bacteria that was swallowed up by the early cells that eventually became all plants and animals. Mitochondria is interesting because it accumulates mutations approximately 10 times the rate of nuclear DNA. Through this fact scientists have traced human DNA back to Africa 200,000 years ago to a single female. Using mitochondrial DNA is helpful in tracing samples of DNA from a long time ago, or a very weak sample, and can be determined easier by the thousands of the mutations.
In this experiment, PCR is used to amplify a 440 nucleotide sequence within the control region of the mt genome. Cycles are much shorter because a sequence is amplified several thousand times with a nuclear sequence with only two copies. Once again we shall be taking cheek cells obtained from our mouths by chewing gently but vigirously. A salt solution is used once again and then the cells are centrifuged and boiled,. The supernatent DNA solution is combined with buffered solution of Taq polymerase, primers, DNTP building blocksl and MgCl2. It is then placed in a thermal cycler

Intro to Mitochondrial DNA

Results/Observations Genetic Disease

This lab was pretty successful looking around the room I saw that only one group had messed up, and this was due to leaving the electrophoresis machine on too long. The results of my table illustrated that everything was done properly, the control illustrated gel electrophoresis worked, the homozygous controls and heterozygous control illustrated the bands that should be seen and were easily comparable to that of mine, Saba's and Daniel's.  The results about the "disease" included the majority of the class as either carriers or diseased. I was personally lucky to be one of the few that did not have the intron, that in reality does not do a whole lot, at least that we know of. I believe this lab was done effectively and I believe reinvigorates me to hopefully take genetic tests in the future in which I could then appropriately assess my risks, and work towards overcoming/ disproving that we are predestined for anything. I hope others understand that the arguement about nature versus nurture is ridiculous, as in everything their is no black or white, we are who we are because of a complex mixture between genetic determination and the society around us including, exposures to environmental sources.