A Summary On ' Pglo Pig Slurry '

70µL of competent E.coli are added to both test tubes; pUC18 and Lux (Alberte et al., 2012). Both test tubes are then tapped and placed back into the ice bath for 15 minutes. While waiting, another test tube is obtained, filled with 35µL of competent cells and labeled NP for no plasmid. A water bath is preheated to 37 degrees Celsius and all three labeled test tubes are inserted into the bath for five minutes (Alberte et al., 2012). Using a sterile pipet 300µL of nutrient broth are inserted into both the control and Lux test tubes and 150µL are inserted to the no plasmid test tube to increase bacterial growth. All three test tubes are then incubated at 37 degrees for 45 minutes. Six agar plates are obtained and labeled to correspond each test tube, three of the plates contain ampicillin. A pipet is used to remove 130µl from each test tube containing a plasmid and insert it into the corresponding agar plate. For this, a cell spreader is first

The plasmid pGLO contains an antibiotic-resistance gene, ampR, and the GFP gene is regulated by the control region of the ara operon. Ampicillin is an antibiotic that kills E. coli, so if E. coli, so if E. coli cells contain the ampicillin-resistance gene, the cells can survive exposure to ampicillin since the ampicillin-resistance gene encodes an enzyme that inactivates the antibiotic. Thus, transformed E. coli cells containing ampicillin-resistance plasmids can easily be selected simply growing the bacteria in the presence of ampicillin-only the transformed cells survive. The ara control region regulates GFP expression by the addition of arabinose, so the GFP gene can be turned on and

This pBlu lab had for purpose to present the changes of the strain of E. coli bacteria due to new genetic information being introduced into the cell. In this experiment we are freezing and heat shocking the E. Coli bacteria that is then forced to take the plasmid DNA. The E. coli then transforms the pBLu plasmid, which carries the genes coding for two identifiable phenotypes. After following the Carolina Biological steps our lab worked well and we able to see some colonies of bacteria on the plates. The x-gal plate showed a significant amount of bacteria to confirm that the pBlu plasmid took over the E. coli strain.

There of the dishes turned out as expected in this experiment. Our group expected there to be growth in the LB -pGLO dish as the bacteria were not exposed to the antibiotic ampicillin. Furthermore, our group also expected to see inhibited bacterial growth in the LB/amp +pGLO dish as there was ampicillin in the dish, but some of the bacteria were immune as they possessed immunity to the ampicillin. Moreover, our group expected that there would be no bacterial growth in the LB/amp -pGLO dish, as the bacteria were exposed to ampicillin and were not immune. However, the final dish, LB/amp/arbo +pGLO, did not turn out as expected. While it was expected to allow for inhibited bacterial growth and the bacteria to become florescent,

Hinton, Page. “The Effect of the Insertion of the pGLO Plasmid on E. coli's” Prezi.com. 10 December 2013Web. 7 Nov. 2016

The purpose of this study was to see whether E. Coli cells would engage in the pGLO plasmid and glow in the presence of four control environmental factors which are arabinose sugar, bacteria, the antibiotic ampicillin, LB nutrient broth and pGLO plasmid DNA. This was tested using four plates, all the plates had E. Coli cells and different environmental factors. The founding was that E. Coli will only fluoresce when bacteria, pGLO plasmid DNA, the antibiotic ampicillin, and LB nutrient broth are present. The result did not support the hypothesis because it stated that, E. coli cells that are exposed to the pGLO plasmid would engage in the plasmid and glow only if the arabinose sugar is present.

If a gene that codes for Green Fluorescent Protein transforms bacteria and GFP glows when transformation occurs, then when two micro test tubes have 250 microliters of transformation solution and places in an ice bath, then 2-4 bacteria colonies are added to each tube with a sterile loop; then a plasmid (pGLO) is added to one of the tubes, incubated in ice for 10 minutes, then heat shocked for 50 seconds at 42 degrees Celsius, then back into 9ice for two minutes; then LB nutrient broth is added to both tubes (250 microliters) and set out at room temperature for 10 minutes. Then, 100 microliters of each solution in the tube are added to four

To start the experiment two small plastic tubes were labeled negative pGLO and positive pGLO as the first step in this experiment. A pipet was then used to move 250 microliters of calcium chloride into each tube and the tubes were iced. Bacteria was added using a new sterilized loop each time to the positive and negative pGLO tubes. The loop was twisted around in the tube to ensure that the bacteria was evenly mixed into the solutions and the tubes were iced once again. Another loop was dipped into the tube containing the plasmid and it was removed when there was a visible slight film of residue. The plasmid was dipped into the tube that was labeled positive for the pGLO gene. The two tubes labeled positive pGLO and negative pGLO were iced

Aim – To test the effectiveness of various anti-microbial substances on the growth of a bacteria called Escherichia coli, also known as E. coli.

Coli. To be able to activate the GFP gene the presence of the sugar arabinose is required. In order to make the E. Coli grow, the presence of ampicillin, an antibiotic, and pGLO is present, however, if ampicillin is present without pGLO it will stop the growth of the E. Coli because the pGLO would not be present to stop the antibiotic. In the experiment E. Coli was tested with the different proteins and plasmids, testing if the plate would glow or grow. If the E. Coli was tested with ampicillin, pGLO, and arabinose the plate would both glow and grow because of the proteins present, however, if the E. Coli was tested with ampicillin and pGLO the plate would neither glow or grow because the proteins were not

This experiment was performed to test the hypothesis if LB nutrient broth, +pGLO and -pGLO Ampicillin, and Arabinose was placed in the E. coli plates, then there will be a significant growth in the newly transformed bacteria and it will possess the ability to glow under UV light. The measurements were recorded from the bent glass tube in each glass test tube. The transformation protocol tested for the newly possessed traits in E.coli bacteria. Throughout the experiment there were many probable reasons for failure. If the pipettes and sterile loop were not thrown out in between each use, a cross contamination could cause a miscalculation in the experiment causing the data results to fail. The hypothesis that was tested was validated due to the positive results with each experiment stating that newly transformed organisms due in fact pass on traits.

It was also conducted to learn about the process of moving genes from one organism to another with the help of a plasmid. The control group was the -pGlo with LB and ampicillin antibiotic, and the -pGlo with LB. The experimental group was the +pGlo with LB and ampicillin antibiotic, and the +pGlo with LB, ampicillin antibiotic, and arabinose. The dependent variable was the bacteria. It had the ability to change. The independent variable is the ampicillin antibiotic and pGlo. They stand alone and are not changed by other factors. The organism used was the bacteria GIVE TYPE OF BAC. and HOW WILL THE BACTERIA BE

The two control plates LB AMP-PGLO and LB AMP+PGLO (refer to Figure 3.0 and Figure 4.0) include a nutrient causing growth and the antibiotic, ampicillin, with or without the plasmid DNA. The purpose of the two is to provide a determination of the LB AMP ARA+PGLO and whether or not, the plasmid has an effect with the sugar. In addition, the two controls show that plasmid, pGLO, can affect the growth of bacteria with the consideration of ampicillin. As well, the two control plates prove that the results of the LB AMP ARA+PGLO and the LB-PGLO plate can be reliable knowing that they have been under the same conditions of the incubator as the control plates. Green Fluorescent Protein acts as the biological molecule for the fluorescence.

Many people think of Escherichia coli as only a severe intestinal illness caused by eating uncooked, contaminated water, or unwashed fruits, but it is much more than only a negative (Amenu et al., 2014). The quick regeneration time of Escherichia coli makes it extremely useful in laboratory studies. In terms of research, a useful aspect of Escherichia coli is the extensive amount of information we have on them (Archer et al., 2011). Scientists have used Escherichia coli to host proteins, and it has been excellent resource for evolution studies as the bacteria has adapted frequently over the years (Archer et al., 2011). The non- pathogenic strains of Escherichia coli are frequently used in medical investigations involving diseases that are difficult to work with. Examples of these diseases include cancer or anthrax poisoning (Das et al., 2013).

The bacteria that was used in the experiment was Escherichia coli, it is one of the most commonly used organisms for gene transfer, and most E. coli are non pathogenic. A non pathogenic strain was used to conduct the experiment. The conditions was kept at 37 ◦C, which is the temperature of the human intestines (BIO-RAD 2010). Arabinose is a sugar that the bacteria needs for energy, used as a food source and enables GFP, which in turn is regulated by araC (Redway 2013). Hypothesis: +pGLO LB/amp will subsist