In this experiment, the bacteria, E Coli, was transformed with the Green Fluorescent Protein (GFP). To start, the bacteria was grown, harvested, and added to a tube with -DNA. Calcium chloride was also added to the tube to allow the cells to take up the DNA and become transformed. Half of this mixture was then placed into a tube with +DNA, which contains the Green Fluorescent Protein, and both were placed in an ice bath for 10 minutes. When time was up, the tubes were placed in 42℃ water for 90 seconds. Next, the tubes were put back in ice water for 2 minutes. Then recovery broth was added to give the bacteria more fluid to grow in and provide nutrients, and the tubes were placed into 37℃ water for 15 minutes. The purpose of placing the tubes in warm and cold water was to heat shock the mixture so the cell membranes would become permeable and the DNA could enter the cells. Once the heat shocking was complete, the -DNA and +DNA were placed on 4 petri dishes with agar. The -DNA was placed on one dish with only agar, and another dish with …show more content…
ampicillin to serve as controls in the experiment. The +DNA was placed on a dish with ampicillin, and another with ampicillin and IPTG. The purpose of using ampicillin was so only the transformed bacteria could grow, because they contain the B-lactamase gene on the plasmid to provide antibiotic resistance, unlike the untransformed bacteria. IPTG was also used to activate the GFP gene in the transformed bacteria. After completing the procedure, the petri dishes were left so the bacteria could grow and then were observed. When looking at the dishes, it was discovered that only petri dish 1, containing only -DNA, had any bacterial growth. There was no bacterial colonies observed on the other three petri dishes, but this is not the outcome that was expected. It was expected to see growth on all the petri dishes, except the dish containing -DNA and ampicillin. Even though three of the four dishes contained ampicillin, which would have limited the growth of the bacteria, there should have been growth seen in the transformed bacteria. Plate 3, with +DNA and ampicillin and plate 4, with +DNA, ampicillin and IPTG, contained transformed bacteria and growth should have occurred here. Plate 1, with only -DNA, and plate 2, with -DNA and ampicillin, did not contain any transformed bacteria, but growth should have occurred on plate 1 because there was no ampicillin present to kill the bacteria. A possible explanation for why these results occurred could be due to decreasing the time the DNA and bacteria was heat shocked. When the tubes were placed in the 37℃ water, they were originally supposed to remain there for 30 minutes, but in this experiment the time was cut in half to 15 minutes. Due to this change, the bacteria may not have been properly heat shocked, affecting the DNA’s ability to enter the cell. If the bacteria didn’t contain the new DNA, then it would not have been equipped with the B-lactamase gene to fight against the ampicillin, and the bacteria would have died. Genetic engineering and bacterial transformation are both beneficial applications in the science field, being used for many purposes.
One example of this technology is the use of bacterial transformation to make insulin. The vector for this process is the E Coli bacteria (Veloso). The gene that codes for insulin comes from human DNA, found on the eleventh chromosomes. This gene is cut using a restriction enzyme, and inserted into a plasmid cut with the same restriction enzyme so the new DNA will fit in the plasmid, which is then inserted into the E Coli bacteria. When the bacteria multiplies, the plasmid is also duplicated with every new bacteria, meaning so is the insulin (Ovsov). Since the bacteria grows and produces insulin, this insulin can be collected for human use. This is very beneficial to people like diabetics, who need insulin to manage and control their blood sugar levels, but may not be able to make enough or
any.
Once the recombinant plasmid was obtained, it was then inserted into E. coli cells through transformation. From a successful transformation, we expected the bacterial cells to translate the inserted EGFP sequence into its protein form. The bacteria cultures were plated on petri dishes containing growth supplement, Luria Broth (LB), an antibiotic: Kanamycin, and IPTG which induced the fluorescence property within successfully transformed bacterial colonies. Different variants of the petri dishes were also included as control and unknown.
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.
...et light. If the LAA plate glows green under exposure to ultraviolet light, then we can conclude that our unknown insert piece of DNA would be the kan gene. If it does not glow green under exposure to ultraviolet light, then then we streak the colony from our LAA plate onto the LAC plate using a sterile glass spreader. When the LAC plate is dray, we place it upside down in the microfuge rack so that it can be incubated at 37 ºC. Incubation at 37 ºC will allow the transformed bacterial cells to grow. If we see bacterial growth on the LA plate containing chloramphenicol, we can conclude that our unknown insert piece of DNA would be the cat gene, since the cat gene is resistant to chloramphenicol. Afterwards, we then grab the microfuge tube labeled NP and repeat the aforementioned steps shown above pertaining to the LA plates. This would be considered our control.
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
What do bacteria need to grow? For bacteria to grow the most typical thing that they like ate a warm and moist environment, but that is not all that they like. Bacteria also like and environment with a PH that is normal or close to a human PH and bacteria also like an oxygen rich environment. The places that could be common to find bacteria in a building are a keyboard, a water fountain, and restrooms. A keyboard is a common place for bacteria because it is being touched constantly with hands when people type and hands are warm, so bacteria like them. The water fountain is another place that is common for bacteria to grow because people's warm hands are touching it and also it has water, which causes it to be moist. The last place that bacteria will we commonly found in buildings are restrooms. The bacteria like restrooms because many people are in then and also there is a lot of water in them.
Eastfield College Microbiology Laboratory Manual, 1st edition, Oliver, T. D. (Book Must Be Purchased New from Eastfield Bookstore and Cannot Be Sold Back to Bookstore at the End of the Semester), Kendall Hunt Publishing, 2013, Dubuque, IA. ISBN 9781465223784.
Genetic engineering is defined as the direct manipulation of genes for practical purposes (citation). It is modifying an organism’s genome using biotechnology (citation). An example of genetic engineering is recombinant DNA technology, which is using DNA from two different sources (citation). This means you can insert the DNA from one species into another in order to make useful proteins. This technology can be used to develop useful human proteins. Some proteins that have been made from recombinant DNA technology are insulin, HGH, Ce...
This line graph shows how some phenotypes were more successful than others. This is an accurate representation of natural selection. The dark blue, pink, and orange phenotypes became extinct before the experiment was even finished. The successful phenotypes were green, purple, and yellow.
In our Biology Lab we did a laboratory experiment on fermentation, alcohol fermentation to be exact. Alcohol fermentation is a type of fermentation that produces the alcohol ethanol and CO2. In the experiment we estimated the rate of alcohol fermentation by measuring the rate of CO2 production. Both glycolysis and fermentation consist of a series of chemical reactions, each of which is catalyzed by a specific enzyme. Two of the tables substituted some of the solution glucose for two different types of solutions. They are as followed, Table #5 substituted glucose for sucrose and Table #6 substituted the glucose for pH4. The equation for alcohol fermentation consists of 6 Carbons 12 Hydrogens 6 Oxygen to produce 2 pyruvates plus 2 ATP then finally the final reaction will be 2 CO2 plus Ethanol. In the class our controlled numbers were at Table #1; their table had 15 mL Glucose, 10 mL RO water, and 10 mL of yeast which then they placed in an incubator at 37 degrees Celsius. We each then measured our own table’s fermentation flasks every 15 mins for an hour to compare to Table #1’s controlled numbers. At
Early examples of genetic engineering include selective breeding and hybridization. Victor Frankenstein’s creation is, in a sense, a hybrid because it is made up of many different pieces intended to yield a superior form, but his plan did not produce the outcome he had wanted. When Frankenstein first lays eyes on his creation, its appearance disgusts him because it is not what he had expected, ‘‘I had selected his features as beautiful. Beautiful! Great God! His yellow skin scarcely covered the work of muscles and arteries beneath; his hair was of a lustrous black, and flowing; his teeth of a pearly whiteness; but these luxuriances only formed a more horrid contrast with his watery eyes, that seemed almost of the same color as the dun-white sockets in which they were set, his shriveled complexion and straight black lips” (Shelley 35). Shelley describes Frankenstein as being utterly disappointed that his carefully thought out process had not brought the outcome he expected. Frankenstein believed that by selecting beautiful pieces the creation would be a beautiful and desirable hybrid, but that is not how the creation ended up being. Instead, it was a disgusting attempt at copying the human form. As the field of research in genetic engineering has grown, new types are being discovered and created. The field has expanded and now includes gene therapy and manipulation of DNA. Gene therapy is divided into two categories. One is the alteration of sperm or egg cells, resulting in a permanent genetic change for the organism and the following generations. The second category is somatic cell therapy, or organ transport (“Genetic Engineering”). Manipulation of DNA is commonly used to alter the genetic makeup of foods to increase the nutrients they contain or to make crops resistant to bugs and
Genetic Engineering is the deliberate alteration of an organism's genetic information (Lee 1). The outcome scientists refer to as successful entitles the living thing’s ability to produce new substances or perform new functions (Lee 1). In the early 1970’s, direct manipulation of the genetic material deoxyribonucleic acid (DNA) became possible and led to the rapid advancement of modern biotechnology (Lee 1).
...ound in the organism. An example would be that organisms are currently on the market and also include plants with the resistance to some insects and also plants can also tolerate herbicides and also crops with modified oil contents. (Biosafety, 2005) Genetic engineering was found to be a changing of an organism. The first big success of genetic engineering was found to be the production of insulin which is a hormone which is produced by the human’s pancreas by something called genetically modified bacteria. Then today’s genetic engineering techniques are used in a lot of different areas.
The Use of Recombinant DNA Technology Recombinant DNA technology is the technology of preparing recombinant DNA in vitro by cutting up DNA molecules and splicing together fragments from more than one organism.(1) This is the process of using recombinant DNA technology to enable the rapid production of human protein from a single gene of insulin. Firstly the single gene required must be isolated. This can be done three ways: Either by working backwards from the protein- Finding the amino acid sequence for the protein needed, the order of bases can be established using known genetic code. New DNA can be made from this sequence of bases resulting in artificial gene made from complementary DNA.
Genetic modification is currently at the forefront of modern science and is being utilised in various fields such as medicine, agriculture and industry. Genetically Modified or transgenic organisms are organisms that have been genetically altered in a specific way for a particular purpose. It is now possible for scientists to exchange genes from one species of organism to another. This process is performed when certain characteristics of one organism are desired in another organism of a different species. For example a pig could be genetically engineered so that it will produce human insulin for those suffering from diabetes. Also, it is seen that it could be possible to cure certain allergies or diseases by replacing the genes responsible for causing the allergy or disease in one organism with that of a gene belonging to an organism that has a resistance to the specific allergen.
Transgenesis has a really high value in producing medicine. In 1974, Cohen transferred the gene of Staphylococcus aureus that against penicillin to colon bacillus, which started humans’ study of transgenesis. Than, in 1982, an American company called Lily successfully reorganized colon bacillus and made it became insulin. Thus, the first medicine produced by genetic engineering appeared. This product was a breakthrough in the history of transgenic technology. Later, Holland foster a transgenic net which had been planted in the human erythropoietin in 1992. Human erythropoientin could stimulate the production of red blood cells and is a useful medicine for healing people who has anemia. With the transgenic technique, the genes of different kinds of organisms could be reorganized, which means humans could reform every organism’s hereditary feature and create a new life style according their own