Plasmid Essays

Result DNA Plasmid Transformation GFP DNA plasmid (Vector Map) Figure 1 Plasmid map of pmaxGFP, encoding the new green fluorescent protein. MaxGFP was used as a positive control for the proper monitoring of transfection efficiency in the HeLa cell. There is comparable transfection efficiencies and mortalities in the HeLa cell that was transfected with MxGFP. HeLa cell transfected with MaxGFP usually reveals slightly higher fluorescent compared to the cells that was expressed with eGFP. This reason

Discussion: Since the plate represented by Figure 1 does not contain plasmid, there will not be any transformed cells. There is also no antibiotic ampicillin, therefore the cells will not die but they will grow and replicate since it is on the LB agar plate. Therefore on this plate there are only the normal E.coli cells. The LB agar plate is a favorable environment for the bacteria to grow and replicate at its optimum rate (Sezonov, 2007). The plate represented by Figure 2 should not have any bacteria

because in the LB broth, we put AMP in there; so the bacteria’s cell are sensitive with AMP. Furthermore, in the LB/AMP positive plate, the bacteria are grow even though the plate contain AMP. However, because bacteria E.coli accepts the pGlo plasmid and the plasmid can encode the gene for resistance to antibiotic, they are survived and grow in single colonies (Nature new, 2017). Finally, in the LB/AMP/Arabinose positive, the bacteria grow with green under the UV light. Like LB/AMP positive plate, the

occurs within plasmids, which are closed circular molecules made up of double stranded DNA. The function of the plasmid is to provide bacteria with genetic advantages such as antibiotic resistance. In this lab, the plasmids provided the ampicillin resistance and the fluorescence. If the bacterial cells are grown in the presence of the antibiotic ampicillin then only the cells that took up the plasmid have the resistance gene. As a result the resistance gene will have to keep the plasmid and the GFP

E. coli Transformation, Plasmid Miniprep and NanoDrop™ Introduction Bacterial transformation is the insertion of DNA into a competent bacterium, which will result in the plasmid expressing the altered gene. A plasmid Miniprep isolates the plasmid DNA by lysing and neutralizing the DNA, followed by a series of washes to get rid of the debris and finally adding an elution buffer to obtain the DNA. A NanoDrop™ determines the absorbance of a DNA sample at 260 nm and its purity at a ratio of ~1.8.

virus that infect bacteria or a plasmid, which is a circular piece of DNA that exist and

n this lab, there were four objectives needed to be met. The first one was to perform a genetic transformation procedure, the second was to move genes from one organism to another using a plasmid as a vector, and the third was to manipulate tools of biotechnology. The bacteria E. coli was used to manipulate and transform. The E. coli would be tested for ampicillin resistance and a green fluorescent glow. One hypothesis made for this lab was that the bacteria that developed a resistance to ampicillin

DNA is inserted into the bacteria's original DNA to alter the genome for a certain outcome. This is usually done by using a plasmid to transfer and incorporate the foreign DNA into the original genome. First, bacterial cells are centrifuged to make a pellet. Then they are shocked with a calcium chloride solution that changes the charge on the cell membrane so that the plasmid DNA may be accepted into the cell. This solution must be chilled so that the cell membrane may heal. After incubating the bacterial

Based off the data that was collected, it is safe to say that if the dish contained the pGLO plasmid, the E. Coli growed. If there was no pGLO in the dish, then the only way E. Coli could grow is if it did not contain ampicillin which is the antibiotic. The only way that the E. Coli could grow is if the dish contained ampicillin and arabinose. Since arabinose is essentially and key that turns on the fluorescent glowing, it is essential to have in order to make the bacteria glow. Since no pGLO was

Bacterial transformation is a technique widely practiced by scientists for research purposes. This experiment explored the transformation of E. coli cultures with pGLO plasmids to allow the bacterial cells to express a foreign protein and emit a fluorescent glow under UV light. The transformation was completed through the heat shock method. Both transformed and untransformed E. coli cultures were grown in four mediums. The four mediums were made of different combinations of the LB nutrient broth

glow. The plate with the plasmid (+pGLO)/LB/amp/ara was the one we said would grow and glow because it contained all the necessary tools to do so. The plate that grew and glowed included the plasmid with the green fluorescent protein (GFP), to make it glow. It also included the resistance gene to ampicillin, as well as the Luria Bertani broth to make the bacteria grow by feeding it, and lastly the arabinose which is in charge of turning on the

too much contamination. The first step in starting the transformation is to add the transformation solution into the +pGLO and -pGLO test tubes. After this is done, you put both tubes in ice and then put bacteria in both tubes. Then, put the pGLO plasmid in the +pGLO test tube but not the -pGLO. Place both of the tubes back in the ice. While the tubes are in the ice, label 4 agar plates: +pGLO LB/amp, +pGLO LB/amp/ara,

Transfection: One of the methods of gene transfer where the genetic material is deliberately introduced into the animal cell in view of studying various functions of proteins and the gene. This mode of gene transfer involves creation of pores on the cell membrane enabling the cell to receive the foreign genetic material. Transfection can be carried out using calcium phosphate (i.e. tricalcium phosphate), by electroporation, by cell squeezing or by mixing a cationic lipid with the material to produce

In this lab, genes for a fluorescent green protein (GFP) and antibacterial resistance (ARG) were inserted into E. coli bacteria. E. coli bacteria was resuspended in an ice-cold CaCl2 solution. DNA containing GFP and ARG was added to half of the cells before they were “heat shocked” in an ice bath and hot water. The heat shocking made the bacteria’s cell membrane more porous, so the DNA could enter. Recovery broth was added to the cell suspension, and the bacteria was placed in warm water for about

different plasmids were used, plasmid lux, and control plasmid, pUC18. The goal for this experiment was to introduce plasmid lux and pUC18 into the bacterium, Escherichia coli, by transformation. (Albert, Pitzer, and Calero et al., 2012). In order for the plasmids to be transformed into E.coli, the E.coli cell has to become a competent cell. By becoming a competent cell, E.coli has the ability to uptake the plasmid DNA, resulting in transformation. The E.coli was then added to both plasmids in two different

restriction map for the unknown plasmid A. The plasmid A was digested with enzymes BAMH1, PstI, and ScaI and then the resulting fragments were run through an agarose gel via electrophoresis. From the gel electrophoresis and deriving an equation by plotting the log of the size of the DNA size markers and distance migrated, a restriction map was constructed. The restriction map showed that the plasmid has only one ScaI site, which supports that vector PRSETB, is present in the plasmid. From the gel electrophoresis

experiment, genetic transformation was successfully carried out. After observing the agar plates, it was found that only the plate with ampicillin and no pGLO plasmid did not grow any of the E.coli bacteria. All three of the other plates grew the E.coli bacteria, however it grew differently in each plate. In the control plate where the pGLO plasmid, ampicillin, and arabinose were not present, the bacteria grew in the pattern that it was spread in originally. In the two other plates, bacteria grew in colonies

inserting a gene into an Escherichia coli bacteria with the help of a plasmid. Escherichia coli bacteria also known as E. coli, is a bacterium that is rod shaped and contains flagella to help it move. The bacterial DNA is circular inside of an E. coli bacterium. E coli. is most known for being found in the intestine of humans and animals but it can also be found in other places such as food

phagocytosis, and this DNA can be either linear or circular, like a plasmid, and is able to either be incorporated into the genome of the bacteria or exist within the cell with degradation. The purpose of using transformation as a laboratory technique is to introduce foreign DNA for many different uses like introducing a specific gene into the bacterial genome that produce a specific protein, medicine, or it can be used to house plasmids in bacteria without the fear of loss during replication. Frederick

used a procedure called heat shock, accompanied by a bacterial plasmid vector, to transform bacteria with a gene that codes for GFP (Green Fluorescent Protein). A vector is an agent “employed to transfer the gene from one organism to another” (Lab manual). Two common vectors are phages and plasmids. Phages are viruses that infect bacteria. Plasmids are “small circular pieces of DNA found within bacteria cells” (Lab manual). Plasmids “often contain genes for traits that are beneficial to the bacteria