An Experiment Conducted in Attempt to Transform DNA off the Bacteria E. Coli

An Experiment Conducted in Attempt to Transform DNA off the Bacteria E. Coli Abstract: The main objective of this experiment was to transform DNA found in bacteria. A plasmid was used on E. coli and with the use of heat shock, was inherited by the bacteria which caused the E. coli to become resistant to ampicillin. Also in the plasmid was GFP which verified the hypothesis by glowing green under UV light. The predicted outcome held true and gives insight on what the future of medicine might hold. Introduction: Transforming DNA on bacteria will require a change in their genes. Using pGLO in this experiment allowed the DNA on Escherichia coli (E. coli) to transform. There are three genes in this plasmid: the araC gene, green fluorescent protein gene (GFP), and the bla gene. The ara C gene is a bifunctional regulator which transcripts araC mRNA which translates to produce araC proteins that act as a depressor or promoter to the GFP. The GFP also transcripts to produce GFP mRNA which translated to produce GFP that glows green under ultraviolet light (UV light). The last gene is the bla gene which gives the bacteria ampicillin resistance. E. Coli was used in this experiment because it is found inside our bodies so it is not very harmful to humans and grows rapidly. Bacteria transformation is the process by which foreign DNA is introduced into a cell (addgene.org). Using plasmids, A linear or circular double-stranded DNA that is capable of replicating independently of the chromosomal DNA(biolog y-online.org), transformation is able to occur. Using heat shock, most of the bacteria accepts the foreign DNA and incorporates it into its own DNA. In this experiment E. coli was transformed to become resistant to the antibiotic ampicillin and expressed the GFP. Using heat shock and pGLO will transform E. coli to be resistant to ampicillin while glowing green. These results could help transform human DNA genes and give resistance to life-threatening diseases such as AIDS. Methods and Materials: Two micro test tubes were labeled +pGLO and the other –pGLO. Using a sterile transfer pipette, 250Â µL of transformation solution (CaCl2) were transferred to each test tube. The test tubes were placed in a bucket of ice for 3 minutes. A single colony of E. coli was placed in the transformation solution (CaCl2) of the test tube labeled +pGLO with a sterile loop and spun until the colony was all the way in the transformation solution. The same was done for the test tube labeled –pGLO. After, both test tubes were placed back in the bucket of ice for another 3 minutes. A new sterile loop was used to transfer pGLO Plasmid DNA to the test tube labeled +pGLO, not –pGLO. The test tubes were then back in the ice bucket for 10 minutes. While the test tubes were in the ice, the agar plates were gathered and labeled with the plate type and group name. After 10 minutes of the test tubes being in ice, they were both transferred to a water bath set at 42Â °C for 50 seconds. After 50 seconds the test tubes were placed back in the ice bucket for 2 minutes. Taking both test tubes out, a sterile pipette was used to pipette 250Â µL of LB nutrient broth into a test tube then mixed well. The same procedure was done to the other test tube using a new sterile pipette. Once both were mixed, they were incubated at room temperature for 20 minutes. After the 20 minutes are done and using a new sterile pipette for each tube, 100Â µL of the transformation and control suspensions were transferred to the corresponding agar plates. Using a new sterile loop for each plate, the suspensions were evenly spread around the surface of the LB nutrient on each plate. The plates were stacked, labeled with a group name, and placed upside down in a fridge at 37Â °C until next week. Independent Variable: Whether or not the plate contained pGLO Dependent Variable: Growth rate Controlled Variables: Amount of LB nutrient Amount of transformation solution Amount of suspensions Time in the ice bucket and room temperature Temperature of water bath and fridge Positive Control: -pGLO, LB Negative Control: -pGLO, LB, AMP These controls were selected because even without the plasmid, the plate labeled ‘-pGLO, LB’ should have growth on it because the only thing added to the plate was nutrients for the bacteria. The plate without the plasmid, LB nutrient, and ampicillin is a negative control because the ampicillin should kill off all the bacteria on the plate. Discussion: The hypothesis was supported in this experiment. In the plate labeled ‘+pGLO, LB, AMP, ARA’ there was growth when normally the ampicillin would kill of the bacteria, as shown in the plate labeled ‘-pGLO, LB, AMP’. Also looking at the UV Light figure, the same plate was able to glow as a cause of inheriting the GFP and being in the presence of arabinose sugar. In the plate labeled ‘-pGLO, LB’ there should be bacterial growth, but there is none present. This could be a possible error in not transferring a big enough colony of E. coli. The plate labeled ‘+pGLO, LB, ARA’ should have more distinct colonies glowing however on the UV Light figure only a film is seen glowing. This could be a cause of putting too much pressure on the colony when spreading the E. coli on the plate. Having the ability to genetically modify DNA could help researchers in the medicine field develop cells that would be resistant to any harmful thing, like the flu or HIV. In conclusion, altering bacteria’s DNA is possible and so easy it can be performed by freshmans in college.

Accounting Essay Example | Topics and Well Written Essays - 500 words - 36

Accounting - Essay Example Failing to provide the cash flow statement, the company will lack the proper information on telling the sources of cash and its use. Additionally, the company will not have adequate planning tool for its business success in the long term (â€Å"Cash flow statements†, 2000). To improve operations, I would compare the expenses that accumulate for a given operation to its budget and provide management warning if the expenses appear to be ahead of projections (Lawrence, 2004). This will give the company time to control cost over the remaining part of the project. I would also approach the client about the increase in billing to cover the cost overrun. Assuming the costs are reimbursed to the clients, I would carry out cost precision where the customer will pay all the expenses incurred plus the revenue. Some of the accounting activities that I would take into consideration will be the cost of materials, cost of labor, and overhead. I would use cost information history to arrive at standard rates and then assign this standard costs to jobs based on the activity units (Drury, 2002). I will allocate the manufacturing overhead cost via Departmental machine hours (Innes & Mitchell, 2003). This is because the manufacturing companies studies and controls the time for direct labour and motion. They have started using machines to replace the direct labor. Using machines increases the factory overhead due to machinery depreciation, machinery maintenance, and set up of machines. Reducing the direct labor and increasing the manufacturing overhead, the correlation between the manufacturing overhead and direct labour tends to wane. Therefore, the logical response is to allocate manufacturing overhead based on the machine hours rather than direct labor hours (Carroll, 2004). I would use break-even analysis to determine the optimum output level below which will

British Airways and its predecessor companies Essay

British Airways and its predecessor companies - Essay Example Two other airlines, Handley page, and Instone, were established using modified bombers. The three companies underwent a period of great difficulty, especially competition from French airlines, which were cheaper. To solve these problems, they merged to be joined later by British Marine Air Navigation, forming Imperial Airways. Imperial Airways began local and overseas flights immediately, flying as far as Egypt and India with a crew of 250 and a fleet of 18 crafts (Gaskell, 2010). This paper is an essay on British Airways. Later, Imperial Airways was a Brisbane, Australia route, whose duration would take grueling 12 days. The new airline added new planes such as the short S.23 C-class model, which signified that the airline was growing, as was a new carrier British Airways limited (Gaskell, 2010). After the start of the 1st World War, these two merged to form British Overseas Airways Corporation, which re-started its transatlantic flights after the war ended. In addition, they create d the BEA, a new airline to handle the European flights. At this point, the carriers needed to order new and more efficient aircraft. BOAC consequently ordered the Boeing Strato-cruiser, the Lockheed Constellation, and a Rolls-Royce engine equipped version of the DC-4. It did not take long before they ordered a jet plane, the De Havilland Comet, which dramatically reduced the length of trans-Atlantic flights (Marriott, 2010). The early 60’s saw BOAC order the Rolls-Royce Conway engine driven 707-436 to tide over until the VC-10s were ready. By 1970, with the first 747 and rapid growth, BOAC and BAL were ready to merge and work as one, establishing British Airways in 1976. BA’s most crucial year was 1976; it had a partnership with Concorde, coupled with big fleets of Lockheed TriStar and Boeing 747.