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DNA as an investigative tool
DNA as an investigative tool
Gel electrophoresis key terms
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The Thermus Aquaticus project’s purpose is to identify a species of thermophilic bacteria found in a local hot spring, Vulcan Hot Springs, through genetic sequencing and analysis. The polymerase gene in other Thermus bacterias has proven useful in genetic reactions. The Vulcan bacteria grows at a higher temperature than other thermophilic bacterias, giving it the potential to have a more effective polymerase gene than what is currently available. My own research has been focused on designing effective primers for the polymerase gene in the bacteria. I have designed primers based on the similarities between the Vulcan bacteria and known Thermus species found in GenBank after growing, isolating, running polymerase chain reaction (PCR), running gel electrophoresis, and sequencing samples in the labs at Cascade High School and Idaho National Laboratories. I have not been able to obtain consistent and correct results from these primers.
INTRODUCTION
Cascade High School’s (in Cascade, Idaho) research lab projects are student taught – passed down from one student to the next each year. Thermus Aquaticus (TAQ) is a research lab project focused on identifying a thermophilic bacterium found in a local hot spring, Vulcan Hot Springs. The polymerase gene in bacteria species, mainly Thermus aquaticus, has proven useful in polymerase chain reactions (PCR), an important reaction in genetic and forensic sciences. The Vulcan bacterium grows at a higher temperature than Thermus aquaticus; the polymerase gene from Vulcan may prove to be more useful than those currently available.
I joined TAQ as a sophomore in high school and learned much of the project, especially the procedures completed on a computer, on my own. After sequencing the 16Sr...
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... extra analysis before ordering primers might have given me more insight into how likely the primers are to work, which may have influenced me to change which primers I ordered. One can always look back at their previous work and wish that he or she could go back in time with their current knowledge and repeat it, but I am pleased with my work and proud that I was able to design primers. I am still hopeful that different PCR settings will change the effectiveness of the primers I designed.
Works Cited
Kennedy, Clinton. Instruction on genetics and reaction processes. 2009-2011.
Miller, Kenneth R. and Joseph S. Levine. “Chapter 12: DNA and RNA.” Biology. Upper Saddle River: Pearson Education, Inc., 2002. Print.
Roberto, Francisco. Instruction on genetics and reaction processes. 2009-2011.
Stocks, Nick. Instruction on genetics and reaction processes. 2009.
...indole, it is motile, there is no urease present and there is no coagulase activity. By deduction and logical reasoning Unknown 10a was determined to be Escherichia coli.
In this lab project, the microbiology students were given 2 unknown bacteria in a mixed broth each broth being numbered. The goal of this project is to determine the species of bacteria in the broth. They had to separate and isolate the bacteria from the mixed broth and ran numerous tests to identify the unknown bacteria. The significance of identifying an unknown bacteria is in a clinical setting. Determining the exact bacteria in order to prescribe the right treatment for the patient. This project is significant for a microbiology students because it gives necessary skills to them for future careers relating to clinical and research work.
They can be found anywhere and identifying them becomes crucial to understanding their characteristics and their effects on other living things, especially humans. Biochemical testing helps us identify the microorganism present with great accuracy. The tests used in this experiment are rudimentary but are fundamental starting points for tests used in medical labs and helps students attain a better understanding of how tests are conducted in a real lab setting. The first step in this process is to use gram-staining technique to narrow down the unknown bacteria into one of the two big domains; gram-negative and gram-positive. Once the gram type is identified, biochemical tests are conducted to narrow down the specific bacterial species.
The purpose of this laboratory is to learn about cultural, morphological, and biochemical characteristics that are used in identifying bacterial isolates. Besides identifying the unknown culture, students also gain an understanding of the process of identification and the techniques and theory behind the process. Experiments such as gram stain, negative stain, endospore and other important tests in identifying unknown bacteria are performed. Various chemical tests were done and the results were carefully determined to identify the unknown bacteria. First session of lab started of by the selection of an unknown bacterium then inoculations of 2 tryptic soy gar (TSA) slants, 1 nutrient broth (TSB), 1 nutrient gelatin deep, 1 motility
The eighteenth exercise of the laboratory manual titled Unknown Identification and Bergey’s Manual is an experiment to identify an unknown bacterium. In this exercise, a student must randomly choose a numbered bacterium available to the class. The keys in Appendix H, located on the last pages of the book, are the major helpful tools in this exercise because it provides completed steps of tests that needs to be performed in order to distinguish certain bacteria. This means that in this exercise, various types of tests and techniques must be performed to identify the chosen unknown bacterium. The unknown bacterium that I selected was number thirty-nine in which I discovered as the Bacillus megaterium after conducting several tests.
small sample in need of identification, Polymerase Chain Reactions are used to multiply the DNA
In the last decade, the number of prescriptions for antibiotics has increases. Even though, antibiotics are helpful, an excess amount of antibiotics can be dangerous. Quite often antibiotics are wrongly prescribed to cure viruses when they are meant to target bacteria. Antibiotics are a type of medicine that is prone to kill microorganisms, or bacteria. By examining the PBS documentary Hunting the Nightmare Bacteria and the article “U.S. government taps GlaxoSmithKline for New Antibiotics” by Ben Hirschler as well as a few other articles can help depict the problem that is of doctors prescribing antibiotics wrongly or excessively, which can led to becoming harmful to the body.
"Polymerase Chain Reaction (PCR) Fact Sheet." National Human Genome Research Institute. 10 Dec. 2007. National Institutes of Health. .
As a process engineer at ZMD, I am conducting this study to identify areas of opportunities for quality improvement. For this purpose, I have taken the initiative in researching the alternative brands to the primer paint currently used. To analysis these brands, I have requested for sample data for three alternatives along with the one currently in use. On the basis of data, my main objective to see which primer including the one that is in use, gives
Since Woese’s research, Archaea have been divided into two main phyla, the Eutyarchota and Crenarchaeota, with the majority being extremophiles. This supports the hypothesis that Eubacteria and Archaea had a thermophilic common ancestor that was able to tolerate the hot conditions on Earth. Nelson et al (1999) also found that Thermotoga maritima bacteria had 24% genes of archaeal origin when analysed, supporting the theory of Thermatoga’s early branching from Archaea in the Tree of Life.
Streptococcus pneumoniae is a Gram-positive and fast-growing bacteria which inhabit upper respiratory tract in humans. Moreover, it is an aerotolerant anaerobe and usually causes respiratory diseases including pneumonia, otitis media, meningitis, peritonitis, paranasal sinusitis, septic arthritis, and osteomyelitis (Todar, 2003). According to Tettelin et al., more than 3 million of children die from meningitis or pneumonia worldwide (2001). S.pneumoniae has an enzyme known as autolysin that is responsible for disintegration and disruption of epithelial cells. Furthermore, S.pneumoniae has many essential virulence factors like capsule which is made up of polysaccharides that avoids complement C3b opsonization of cells by phagocytes. Many vaccines contain different capsular antigens which were isolated from various strains (Todar, 2003). There are plenty of S.pneumoniae strains that developed resistance to most popular antibiotics like macrolides, fluoroquinolones, and penicillin since 1990 (Tettelin et al., 2001). Antibiotic resistance was developed by the gene mutation and selection processes that, as a consequence, lead to the formation of penicillin-binding proteins, etc. (Todar, 2003).
When observed under the microscope, the bacteria’s morphology displayed a purple color with rod shape. This bacteria is found in soil and vegetation. It is most favorable in mesophilic temperature ranging between 25-35 degrees Celsius4
The history of Ab starts when “the Dutch microbiologist, Beijerinck, first isolated the bacteria from soil using minimal media enriched with calcium acetate, in 1911” (Aoife et al. 244). From there different characteristic of Ab were noted, such as, “Ab is often found in pairs, does not use oxygen, does not produce nitrites, does not move, and it can be isolated on a MacConkey agar, which is a specific media of petri dish” (Rosenbaum et al. 10). Using a staining method, it is determined that Ab is a gram-negative bacterium, because it does take on the stain. Although, Ab can be difficult to de-stain, making it harder to distinguish between gram-negative or positive (Aoife et al 244). “It also has the ability to survive longer in any environment, even on dry surfaces, because it is resistance to desiccation, or being completely deprived
Raven, P. H., Mason, K. A., Losos, J. B., Singer, S. R., & Johnson, G. B. (2011). Biology (9thth ed., pp. 418-419). New York City, NY: McGraw Hill.
Different types of bacteria have different range of temperature they are able to survive. They are generally divided into three types: psychrophiles, mesophiles and thermophiles. Psychrophilic bacteria are able to survive in low temperatures ranging from about -10 to 20°C while thermophilic bacteria are able to thrive in high temperatures ranging from 40 to 75°C. These two types of bacteria are also known as extremophiles due to their ability to survive in extreme conditions. Mesophilic bacteria are bacteria that thrive in temperatures ranging from 10 to 45°C and usually have an optimum growth temperature of about 37°C (M. Furlong, n.d.).